Heterocyclic compound as syk inhibitor and/or syk-hdac dual inhibitor

ABSTRACT

A heterocyclic compound as a Syk inhibitor and/or a Syk-HDAC dual inhibitor, or pharmaceutically acceptable salts, prodrugs, deuterated derivatives, hydrates, and solvates thereof are provided. Specifically, a compound of formula (I) is provided, which has dual inhibitory activity for Syk and/or Syk-HDAC.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a divisional of U.S. application Ser. No. 16/468,902filed Jun. 12, 2019, which is a Section 371 of International ApplicationNo. PCT/CN2017/115755, filed Dec. 12, 2017, which was published in theChinese language on Jun. 21, 2018, under International Publication No.WO 2018/108083 A1, which claims priority under 35 U.S.C. § 119(b) toChinese Application No. 201611141394.8, filed Dec. 12, 2016, thedisclosures of which are incorporated herein by reference in theirentirety.

FIELD OF THE INVENTION

The present invention provides a novel class of heterocyclic compounds,the synthesis and use thereof, for example, as Syk (Spleen tyrosinekinase) inhibitors and/or Syk-HDAC (Histone deacetylase) dualinhibitors.

BACKGROUND OF THE INVENTION

Syk is a non-receptor protein tyrosine kinase which is expressed in avariety of cells, especially in various hematopoietic cells. It isexpressed in monocytes, macrophages, mast cells, basophils, eosinophils,neutrophils, immature T cells, CD4 effector T cells, B cells, naturalkiller cells, dendritic cells, platelets and red blood cells. Inaddition, expression of Syk is detected also in fibroblasts,osteoclasts, endothelial cells, and nerve cells. Syk also presents invarious tissues such as epithelial cells of the lung, kidney, andcardiomyocytes. In 1991, Taniguchi et al. isolated a protein kinase witha molecular weight of 72 kDa from the cDNA of pig spleen, and named itSyk. Syk contains 629 amino acid residues and consists of two tandem Srchomodomains (N—SH2 and C—SH2) at the N-terminus and a kinase domain atthe C-terminal. It shares a part of common structure with the proteinkinase (ZAP-70), which is also a cytoplasmic protein kinase. Syk isactivated by binding of the SH2 region to a tyrosine-dependentimmunoreceptor tyrosinebased activation motifs (ITAM).

Spleen tyrosine kinase is involved in the signal transduction process ofmany cells, and has attracted extensive attention as a cell signaltransduction factor, especially an immune signal transduction factor.Recent studies have shown that Syk plays a key role in inhibiting celldivision and proliferation, and its overactivation can promote malignantcell proliferation and inhibit apoptosis, especially in B cells. Sykalso affects the secretion of certain cytokines, and plays a key role inthe production of cytokines in T cells and monocytes, bone resorption inosteoclasts, and phagocytosis of macrophages. Syk also affects thematuration and activation of immune cells and is closely related toallergic and antibody-mediated autoimmune diseases. Since Syk is locatedin the upstream of the cellular signaling pathway, treatments targetingto Syk are more advantageous than drugs that inhibit single downstreampathway. Therefore, Syk has been used as a potential therapeutic targetfor a variety of diseases, such as chronic inflammatory diseases such asrheumatoid arthritis, allergic diseases (allergic rhinitis and asthma),multiple sclerosis, and immune diseases (rheumatoid arthritis), multipletumors (breast, stomach, rectal, pancreatic, liver, B-cell lymphoma,chronic lymphocytic leukemia, non-Hawkings lymphoma, etc.),atherosclerosis (coronary heart disease and cerebral arterialthrombosis), gastrointestinal disorders, idiopathic thrombocytopenicpurpura, Wiskott-Aldrich syndrome, and systemic lupus erythematosus.

HDAC is a class of proteases that play an important role in thestructural modification of chromosomes and the regulation of geneexpression. HDAC deacetylates the lysine side chain at the aminoterminus of histones, and histone acetylation is in a dynamicequilibrium with histone deacetylation, which is regulated by histoneacetyltransferase (HAT) and histone deacetylase. The acetylation ofhistones reverses the acetylation of lysine residues of HAT and restoresthe positive charge of lysine residues, which facilitates thedissociation of DNA and histone octamers, and the relaxation ofnucleosome structures, thus making various Transcription factors andco-transcription factors bind specifically to the DNA binding site andactivate transcription of the gene. Due to the overexpression of HDAC intumor cells, the deacetylation of histones is enhanced. By restoring thepositive charge of histones and increasing the gravitation between DNAand histones, the relaxed nucleosomes become very tight, which is notconducive to specific Gene expression, including some tumor suppressorgenes.

HDAC inhibitors can regulate the expression and stability of apoptosisand differentiation-related proteins by increasing histone acetylationin specific regions of chromatin, induce tumor cell cycle arrest andapoptosis, promote tumor cell autophagy, and inhibit the formation oftumor angiogenesis, promotes the immunogenicity of tumor cells. HDACinhibitors not only become a target for therapy for tumors, but alsoplay a role in neurological diseases, inflammation, and promotion ofautoimmunity.

Preclinical and clinical studies have shown that HDAC inhibitors canalso effectively synergistically inhibit tumor growth when combined withother anti-tumor compounds. HDAC is responsible for removing acetylgroups on histones, which shows significant effects on gene expression,oncoprotein stability, cell migration, protein catabolism, and cellcycle regulation.

Study of Hagiwara, K. et al. on 2015 published in Apoptosis confirmedthat the combination of the Syk inhibitor R406 and the HDAC inhibitorvorinostat has synergistically potentiating effects on killing mantlecell lymphoma cells.

In summary, Syk inhibitors or Syk-HDAC dual inhibitors would be able toused in a variety of cancers and other treatments for the disease.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a structurally novelSyk inhibitors or Syk-HDAC dual inhibitors, and the preparation methodand application thereof.

In the first aspect of the present invention, a compound of formula (I),or the pharmaceutically acceptable salts, prodrugs, deuteratedderivatives, hydrates, or solvates thereof is provided:

wherein in the formula (I),

R¹ is aryl, heteroaryl or 6-membered monocyclic heterocyclyl (includingsaturated and unsaturated); aryl, heteroaryl or monocyclic heterocyclylherein may be optionally and independently substituted by 1-3substituents each independently selected from the group consisting of:halogen, C₁₋₄ alkyl, C₁₋₄ halogenated alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl,C₃₋₈ cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN,NO₂, OR⁸, SR⁸, NR⁸R⁹, C(O)R⁸, C(O)OR⁸, C(O)NR⁸R⁹, S(O)₂R⁸ or R⁷.

R², R³ and R⁴ are hydrogen;

U is selected from NR⁵; where R⁵ is hydrogen;

A selected from the group consisting of formula (II) or formula (III):

wherein:

“

” refers to the connection point of formula (II) or formula (III) to Uof the formula (I);

“*” indicates a chiral center;

B is a monocyclic aryl or bicyclic aryl group, or a monocyclicheteroaryl or bicyclic heteroaryl group, and at least one ring of thebicyclic aryl or bicyclic heteroaryl is aromatic, and the other ring isaromatic, saturated or partially saturated ring;

Y is 3- to 12-membered monocyclic or polycyclic heterocyclic ring;wherein said heterocyclic ring contains 1-4 heteroatoms eachindependently selected from N, O and S;

m is 0 or 1;

each X is independently hydrogen, halogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl,3- to 6-membered heterocyclyl, CN, OR⁸, SR⁸, NR⁸R⁹, C(O)R⁸, C(O)OR⁸,C(O)NR⁸R⁹, or S(O)₂R⁸;

R is hydrogen, —(CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH),—V¹—(CH₂)_(p)—V²—V—(CH₂)_(q)C(O)NH(OH);

R⁷ is hydrogen, —(CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH),—V¹—(CH₂)_(p)—V²—V—(CH₂)_(q)C(O)NH(OH), C(O)NH(OCH₃),

J is O;

G is NR¹⁰;

n is 0, 1, 2, or 3;

each R⁶ is hydrogen, or two R⁶ connecting to the same carbon atom formcarbonyl group (═O);

R⁸ and R⁹ are each independently hydrogen, C₁₋₄ alkyl, C₁₋₄ halogenatedalkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₈ cycloalkyl, 3- to 8-memberedheterocyclyl, aryl, or heteroaryl; or R⁸ and R⁹ together with thenitrogen atom to which they are attached form a 3- to 9-metacyclic ringcomprising 1-2 N atom and 0, 1 or 2 heteroatoms selected from O or S;

R¹⁰ is C₂₋₈ alkyl, C₁₋₈ haloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₈cycloalkyl, 3- to 12-membered heterocyclyl (optionally comprising 1-2heteroatoms selected from O, N, or S), aryl, heteroaryl, C(O)R⁸,C(O)OR⁸, C(O)NR⁸R⁹, S(O)₂R⁸ or (CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH), whereinR⁸ is not hydrogen, and the R⁸ in C(O)R⁸ is not methyl;

V is a divalent group, each of p and q is independently an integer from0 to 10, and the V is selected from the group consisting of bond, O, S,NR¹¹, OC(O), OC(O)O, NHC(O), NHC(O)NH, NHC(O)O, OC(O)NH, NHS(O)₂, C(O),C(O)O, C(O)NH, S(O), S(O)₂, S(O)₂NH, or NHS(O)₂NH, CH═CH, C≡C, CR¹²R¹³,C₃₋₈ cycloalkyl, 3- to 12-member heterocyclyl, aryl or heteroaryl,

with the prerequisite that V, p and q together form a chemically stablegroup;

V¹ and V² are divalent groups selected from the group consisting ofbond, O, S, NR¹¹, or C(O)NH, with the prerequisite that the group formedby V, V¹, V², p and q is chemically stable group;

R¹¹ is hydrogen, C₁₋₄ alkyl, C₃₋₈ cycloalkyl, 3- to 8-memberedheterocyclyl, aryl, heteroaryl, C(O)R⁸ or S(O)₂R⁸;

R¹² and R¹³ are each independently hydrogen, halogen, C₁₋₄ alkyl, C₃₋₆cycloalkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, 3- to 8-membered heterocyclic,OR⁸, SR⁸, NR⁸R⁹, CN, C(O)R⁸, C(O)OR⁸, C(O)NR⁸R⁹, OC(O)R⁸, NR⁸C(O)R⁹, orS(O)₂R⁸, or R¹² and R¹³ together with the carbon atoms to which they areattached form 3-8 membered cyclic structure containing 0, 1 or 2heteroatoms selected from N, O, or S;

with the prerequisite that when the formula (II) is

then the R¹ is

herein R⁷ is other than hydrogen, and the definitions of V, V¹, V², pand q in R⁷ must ensure that the formed R¹ group is a stable chemicalstructure;

another proviso is that when R¹ does not comprise structural unit

than A is of formula (IIa) or formula (IIIa), wherein R comprisesstructural unit

wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl,aryl and heteroaryl are each optionally and independently substituted by1-3 substituents each independently selected from the group consistingof halogen, C₁₋₄ alkyl, C₁₋₄ halogenated alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₈ cycloalkyl, 3- to 8-heterocyclyl, aryl, heteroaryl, CN,NO₂, OR⁸, SR⁸, NR⁸R⁹, C(O)R⁸, C(O)OR⁸, C(O)NR⁸R⁹ or S(O)₂R⁸;

unless otherwise specified, the above aryl group is aryl group having 6to 12 carbon atoms; and the heteroaryl group is 5- to 15-memberedheteroaryl group.

In another preferred embodiment, the R¹ is

wherein the R⁷ is as defined above.

In another preferred embodiment, Y is a 4- to 12-membered monocyclic orpolycyclic heterocycle having 1-4 heteroatoms each independentlyselected from N, O or S.

In another preferred embodiment, Y is a 4- to 10-membered monocyclic orpolycyclic heterocycle having 1-3 heteroatoms each independentlyselected from N, O or S.

In another preferred embodiment, B is phenyl; Y is 6-membered monocyclicheterocyclic ring having 1-2 heteroatoms each independently being N, Oor S, or Y is absent (m is 0).

In another preferred embodiment, formula (II) is a structure selectedfrom the group consisting of:

where D is N or CH;

k¹ and k² are each independently 0, 1, 2, or 3;

X is hydrogen, halogen, C₁₋₄ alkyl, CN, or OR⁵;

R is hydrogen, —(CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH),—V¹—(CH₂)_(p)—V²—V—(CH₂)_(q)C(O)NH(OH).

In another preferred embodiment, the formula (II) is a structureselected from the group consisting of:

wherein X is hydrogen, halogen, C₁₋₄ alkyl, CN, or OR⁵; R is asdescribed above.

In another preferred embodiment, the formula (III) is a structureselected from the group consisting of:

wherein R¹⁰ is C₂₋₈ alkyl, C₁₋₈ halogenated alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₈ cycloalkyl, 6-membered heterocyclyl (optionally comprising1-2 heteroatoms selected from O, N, S), aryl, heteroaryl, C(O)R⁸,C(O)OR⁸, C(O)NR⁸R⁹, S(O)₂R⁸, wherein R⁸ is selected from the groupconsisting of C₁₋₄ alkyl, C₁₋₄ haloalkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl,C₃₋₈ cycloalkyl, 3- to 8-membered heterocyclyl, aryl, or heteroaryl;wherein R⁸ in C(O)R⁸ is other than methyl; or R⁸ and R⁹ together withthe nitrogen atom to which they are attached form a 3- to 9-memberedring comprising 1-2 N atom and 0, 1 or 2 hetero atoms selected from O orS.

In another preferred embodiment, R is —(CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH)or —V¹—(CH₂)_(p)—V²—V—(CH₂)_(q)C(O)NH(OH); wherein V is bond, O, NR¹¹,CH═CH, aryl, heteroaryl, OC(O), NHC(O), C(O), S(O)₂; each R¹¹ isindependently hydrogen or C₁₋₄ alkyl; p is 0, 1, 2, 3, or 4; q is 0, 1,2, 3, 4, 5, 6, 7, or 8; V¹ and V² are each independently selected fromNR¹¹, O, S, or bond; with the proviso that the group formed by V, V¹,and V², p and q together is a stable chemical structure.

In another preferred embodiment, each of R¹, R², R³, R⁴, U, A are thecorresponding groups in compounds of the specific formula (I) preparedin the examples, respectively.

In another preferred embodiment, the formula (I) is selected from thegroup consisting of

wherein R¹⁰ is C₂₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈ cycloalkyl,unsubstituted or C₁₋₄ alkyl substituted 6-membered heterocyclyl(optionally containing 1-2 heteroatoms selected from O, or N), C(O)R⁸,or S(O)₂R⁸; R⁸ is C₁₋₄ alkyl or C₁₋₄ haloalkyl; wherein the R⁸ in C(O)R⁸is other than methyl.

In another preferred embodiment, the R¹⁰ is selected from the groupconsisting of ethyl, haloethyl, cyclopropyl, phenyl, 6-heterocyclylunsubstituted or substituted by C₁₋₄ alkyl (optionally containing 1-2heteroatoms selected from O or N), 5-6 membered heteroaryl unsubstitutedor substituted by C₁₋₄ alkyl (optionally containing 1-2 hetero atomsselected from O or N), C(O)R⁸, S(O)₂R⁸; wherein R⁸ is C₁₋₄ alkyl or C₁₋₄halogenated alkyl; wherein the R⁸ in C(O)R⁸ is other than methyl.

In another preferred embodiment, formula (I) is a structure selectedfrom the group consisting of:

wherein V, V¹, V², p, and q are as defined above.

In another preferred embodiment, the formula (I) compound is a structureselected from the group consisting of:

wherein R⁷ is —(CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH) or—V¹—(CH₂)_(p)—V²—V—(CH₂)_(q)C(O)NH(OH); wherein V is bond, O, NR¹¹,CH═CH, aryl, heteroaryl, OC(O), NHC(O), C(O), S(O)₂; each R¹¹ isindependently hydrogen or C₁₋₄ alkyl; p is 0, 1, 2, 3, or 4; q is 0, 1,2, 3, 4, 5, 6, 7, or 8; V¹ and V² are each independently selected fromNR¹¹, O, S, or bond; with the proviso that the group formed by V, V¹,and V², p and q together is a stable chemical structure.

In another preferred embodiment, the compound is selected from thefollowing group:

In the second aspect of the present invention, a use of compound offormula (I) according to the first aspect of the invention, or opticalisomers, pharmaceutically acceptable salts, prodrugs, deuteratedderivatives, hydrates, or solvates thereof is provided, wherein in:

(a) preparation of medicine for treating diseases associated with Sykand/or HDAC kinase activity or expression level;

(b) preparation of Syk and/or HDAC kinase targeting inhibitor; and/or

(c) in vitro non-therapeutic inhibition of Syk and/or HDAC kinaseactivity;

In another preferred embodiment, the formula (I) compound is used totreat diseases associated with Syk and/or HDAC kinase activity orexpression level.

In another preferred embodiment, a method of treating a diseaseassociated with Syk and/or HDAC kinase activity or expression level isprovided, comprising the steps: administering a therapeuticallyeffective amount of a compound of formula (I) to a subject in need.

In the third aspect of the present invention, a pharmaceuticalcomposition is provided, which comprises: (i) therapeutically effectiveamount of formula (I) compound of the first aspect of the invention, oroptical isomers, pharmaceutically acceptable salts, prodrugs, deuteratedderivatives, hydrates, or solvates thereof, and (ii) pharmaceuticallyacceptable carrier.

In the fourth aspect of the present invention, the preparation method ofcompound of the first aspect of the present invention is provided, whichcomprises the following steps:

(1) In an inert solvent, compound Ia reacts with A-NH₂ so as to providecompound Ib;

(2) In an inert solvent, compound Ib reacts with compound R¹B(OH)₂ toobtain formula I compound;

(3) The C(O)NH(OH) group in A or R¹ in the compound of formula (I) isprepared from the corresponding carboxylic ester. The carboxylic esterIc or Id is hydrolyzed, and the resulting acid is further reacted with atetrahydropyran protected hydroxylamine, and finally the tetrahydropyranprotecting group is deprotected to give a hydroxyamide 1e or 1f. Thegeneral scheme is as follows:

In the above formulas, the groups are defined as above.

It should be understood that, in the present invention, each of thetechnical features specifically described above and below (such as thosein the Examples) can be combined with each other, thereby constitutingnew or preferred technical solutions which need not be specified againherein.

EMBODIMENTS FOR CARRYING OUT THE INVENTION

After long-term and intensive research, the inventors have unexpectedlydiscovered a class of heterocyclic compounds having Syk (spleen tyrosinekinase) inhibitory activity or Syk-HDAC dual inhibitory activity, thusbeing able to used in the preparation of pharmaceutical compositions fortreating diseases associated with the activity or expression level ofSyk and or HDAC. The present invention is completed on this basis.

Terminology

Unless otherwise stated, “or” as used herein has the same meaning as“and/or” (refers to “or” and “and”).

Unless otherwise specified, among all compounds of the presentinvention, each chiral carbon atom (chiral center) may optionally be inthe R configuration or the S configuration, or a mixture of the Rconfiguration and the S configuration.

As used herein, the term “alkyl”, alone or as part of anothersubstituent, refers to a straight (ie, unbranched) or branched saturatedhydrocarbon group containing only carbon atoms, or a combination ofstraight and branched chains. When the alkyl group has a carbon numberlimitation (e.g., C₁₋₁₀), it means that the alkyl group has 1 to 10carbon atoms. For example, C₁₋₈ alkyl refers to an alkyl groupcontaining from 1 to 8 carbon atoms, including methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like. Group.

As used herein, the term “alkenyl”, when used alone or as part ofanother substituent, refers to a straight or branched, carbon chaingroup having at least one carbon-carbon double bond. Alkenyl groups canbe substituted or unsubstituted. When the alkenyl group has a carbonnumber limit (e.g., C₂₋₈), it means that the alkenyl group has 2-8carbon atoms. For example, C₂₋₈ alkenyl refers to alkenyl groups having2-8 carbon atoms, including ethenyl, propenyl, 1,2-butenyl, 2,3-butenyl,butadienyl, or the like.

As used herein, the term “alkynyl”, when used alone or as part ofanother substituent, refers to an aliphatic hydrocarbon group having atleast one carbon-carbon triple bond. The alkynyl group can be straightor branched, or a combination thereof. When the alkynyl group has acarbon number limitation (e.g., C₂₋₈ alkynyl group), it means that thealkynyl group has 2 to 8 carbon atoms. For example, the term “C₂₋₈alkynyl” refers to a straight or branched alkynyl group having 2-8carbon atoms, including ethynyl, propynyl, isopropynyl, butynyl,isobutynyl, secondary Butynyl, tert-butynyl, or the like.

As used herein, when used alone or as part of another substituent, theterm “cycloalkyl” refers to a unit ring having a saturated or partiallysaturated ring, a bicyclic or polycyclic (fused ring, bridged or spiro)ring system. When a certain cycloalkyl group has a carbon numberlimitation (e.g., C₃₋₁₀), it means that the cycloalkyl group has 3 to 10carbon atoms. In some preferred embodiments, the term “C₃₋₈ cycloalkyl”refers to a saturated or partially saturated monocyclic or bicyclicalkyl group having from 3 to 8 carbon atoms, including cyclopropyl,cyclobutyl, cyclopentyl, cycloheptyl, or the like. “Spirocycloalkyl”refers to a bicyclic or polycyclic group that shares a carbon atom(called a spiro atom) between the monocyclic rings. These may containone or more double bonds, but none of the rings have fully conjugated 1L electrons system. “Fused cycloalkyl” means an all-carbon bicyclic orpolycyclic group in which each ring of the system shares an adjacentpair of carbon atoms with other rings in the system, wherein one or moreof the rings may contain one or more double bond, but none of the ringshave a fully conjugated R-electron system. “Bridged cycloalkyl” refersto an all-carbon polycyclic group in which two rings share two carbonatoms that are not directly bonded, which may contain one or more doublebonds, but none of the rings have a fully conjugated pi-electron system.The atoms contained in the cycloalkyl group are all carbon atoms. Someexamples of cycloalkyl groups are as follows, and the present inventionis not limited to the following cycloalkyl groups.

Unless otherwise stated, the following terms used in the specificationand claims have the following meanings. “Aryl” means an all-carbonmonocyclic or fused polycyclic (ie, a ring that shares a pair ofadjacent carbon atoms) groups having a conjugated R-electron system,such as phenyl and naphthyl. The aryl ring may be fused to other cyclicgroups (including saturated and unsaturated rings), but may not containheteroatoms such as nitrogen, oxygen, or sulfur, while the point ofattachment to the parent must be on the ring of carbon atoms ofconjugated pi-electron system. The aryl group can be substituted orunsubstituted. The following are some examples of aryl groups, and thepresent invention is not limited to the aryl groups described below.

“Heteroaryl” refers to a heteroaromatic group containing one to moreheteroatoms. The heteroatoms referred to herein include oxygen, sulfur,and nitrogen. For example, furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl,N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and thelike. The heteroaryl ring may be fused to an aryl, heterocyclic orcycloalkyl ring wherein the ring to which the parent structure isattached is a heteroaryl ring. The heteroaryl group can be optionallysubstituted or unsubstituted. The following are some examples ofheteroaryl groups, and the present invention is not limited to thefollowing heteroaryl groups. Among them, the last three heteroarylgroups are tricyclic heteroaryl groups, which are the focus of thepresent invention.

“Heterocyclyl” means a saturated or partially unsaturated monocyclic orpolycyclic cyclic hydrocarbon substituent wherein one or more of thering atoms are selected from nitrogen, oxygen or sulfur and theremaining ring atoms are carbon. Non-limiting examples of monocyclicheterocyclic groups include pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, homopiperazinyl. Polycyclic heterocyclicgroup refers to a heterocyclic group including a spiro ring, a fusedring, and a bridged ring. “Spirocyclic heterocyclyl” refers to apolycyclic heterocyclic group in which each ring of the system shares anatom (referred to as a spiro atom) with other rings in the system,wherein one or more of the ring atoms is selected from the groupconsisting of nitrogen and oxygen. Or sulfur, the remaining ring atomsare carbon. “Fused ring heterocyclyl” refers to a polycyclicheterocyclic group in which each ring of the system shares an adjacentpair of atoms with other rings in the system, and one or more rings maycontain one or more double bonds, but none One ring has a fullyconjugated pi-electron system, and wherein one or more ring atoms areselected from nitrogen, oxygen or sulfur, and the remaining ring atomsare carbon. “Bridged heterocyclyl” refers to a polycyclic heterocyclicgroup in which any two rings share two atoms which are not directlybonded, these may contain one or more double bonds, but none of therings have a fully conjugated pi-electron system And wherein one or moreof the ring atoms are selected from nitrogen, oxygen or sulfur, and theremaining ring atoms are carbon. If a heterocyclic group has both asaturated ring and an aromatic ring (for example, the saturated ring andthe aromatic ring are fused together), the point attached to the parentmust be on the saturated ring. Note: When the point attached to theparent is on the aromatic ring, it is called a heteroaryl group and isnot called a heterocyclic group. Some examples of the heterocyclic groupare as follows, and the present invention is not limited to thefollowing heterocyclic group.

As used herein, the term “halogen”, when used alone or as part ofanother substituent, refers to F, Cl, Br, and I.

As used herein, the term “substituted” (when with or without“optionally”) means that one or more hydrogen atoms on a particulargroup are replaced by a particular substituent. Particular substituentsare the substituents described above in the corresponding paragraphs, orthe substituents which appear in the examples. Unless otherwise stated,an optionally substituted group may have a substituent selected from aparticular group at any substitutable position of the group, and thesubstituents may be the same or different at each position. A cyclicsubstituent, such as a heterocyclic group, may be attached to anotherring, such as a cycloalkyl group, to form a spirobicyclic ring system,i.e., the two rings have a common carbon atom. Those skilled in the artwill appreciate that the combinations of substituents contemplated bythe present invention are those that are stable or chemicallyachievable. The substituents are, for example but not limited to, C₁₋₈alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₈ cycloalkyl, 3- to 12-memberedheterocyclic, aryl, Heteroaryl, halogen, hydroxy, carboxy (—COOH), C₁₋₈aldehyde, C₂₋₁₀ acyl, C₂₋₁₀ ester, amino.

For convenience and in accordance with conventional understanding, theterm “optionally substituted” or “optionally substituted” applies onlyto sites which are capable of being substituted by a substituent, anddoes not include those which are not chemically achievable.

As used herein, unless otherwise specified, the term “pharmaceuticallyacceptable salt” refers to a salt that is suitable for contact with thetissue of a subject (eg, a human) without causing unpleasant sideeffects. In some embodiments, a pharmaceutically acceptable salt of acompound of the invention includes a salt (eg, a potassium salt, asodium salt, a magnesium salt, a calcium salt) of a compound of theinvention having an acidic group or is basic A salt of a compound of theinvention (e.g., a sulfate, a hydrochloride, a phosphate, a nitrate, acarbonate).

General Synthetic Method of Compounds

The compound of the formula (I) of the present invention can be preparedby the following method:

(1) in an inert solvent, reacting formula Ia compound with A-NH₂ so asto provide compound Ib;

(2) In an inert solvent, reacting compound Ib and R¹B(OH)₂ compound toobtain formula I compound;

In the above formulas, the groups are defined as above. The reagents andconditions for each step may be selected from those conventional in theart for carrying out such preparation methods. After the structure ofthe compound of the present invention is disclosed, the above selectionmay be carried out by those skilled in the art based on the knowledge inthe art.

More specifically, the compound of the formula I of the presentinvention can be obtained by the following method, however, theconditions of the method, such as the reactant, the solvent, the base,the amount of the compound used, the reaction temperature, the reactiontime, are not limited to the explanation below. The compounds of theinvention may also be easily prepared by optionally combine varioussynthetic methods described in this specification or known in the art,such a combination can be easily performed by one of ordinary skill inthe art of the present invention.

In the production method of the present invention, each reaction isusually carried out in an inert solvent, and the reaction temperature isusually −20 to 150° C. (preferably 0 to 120° C.). The reaction time ofeach step is usually 0.5 to 48 h, preferably 2 to 12 h.

General Synthesis Method:

Scheme A describes the general synthesis method of the compounds A9-1and A9-2:

Scheme B describes the general synthesis method of the compound B4:

Scheme C describes the general synthesis method of the compound C9:

Scheme D describes the general synthesis method of the compound D5:

Pharmaceutical Composition and Method of Administration

Since the compound of the present invention has excellent Syk kinasesinhibitory activity and/or Syk-HDAC dual inhibitory activity, thecompound of the present invention, the related deuterated compoundsthereof, the possible isomers thereof (enantiomers or diastereoisomersthereof), and various crystal forms thereof, pharmaceutically acceptableinorganic or organic salts, hydrates or solvates, and pharmaceuticalcompositions comprising compounds containing the present invention canbe used to treat, prevent, and alleviate diseases associated with Syk orHDAC activity or expression levels. According to the prior arts, thecompound of the present invention can be used to treat diseases selectedfrom the group consisting (but not limited to) lymphoma, lymphocyticleukemia, cutaneous T-cell lymphoma, rectal cancer, breast cancer,stomach cancer, pancreatic cancer, liver cancer, lung cancer, head andneck cancer, kidney cancer, colon cancer, ovarian cancer, prostatecancer, multiple sclerosis, immunity diseases (rheumatoid arthritis andnephritis), allergic diseases (allergic rhinitis and asthma),atherosclerosis (coronary heart disease and ischemic stroke),gastrointestinal disorders, idiopathic thrombocytopenic purpura,systemic lupus erythematosus, Alzheimer's disease, stroke and coronaryartery disease, Wiskott-Aldrich syndrome, myelofibrosis, AIDS, etc.

The pharmaceutical compositions of the present invention comprise a safeor effective amount of a compound of the present invention, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient or carrier. By “safe and effective amount” it ismeant that the amount of the compound is sufficient to significantlyimprove the condition without causing serious side effects. In general,the pharmaceutical compositions contain from 1 to 2000 mg of thecompound of the invention per agent, more preferably from 5 to 200 mg ofthe compound of the invention per agent. Preferably, the “one dose” is acapsule or tablet.

“Pharmaceutically acceptable carrier” means: one or more compatiblesolid or liquid fillers or gel materials which are suitable for humanuse and which must be of sufficient purity and of sufficiently lowtoxicity. By “compatibility” it is meant herein that the components ofthe composition are capable of intermingling with the compounds of theinvention and with each other without significantly reducing theefficacy of the compound. Examples of pharmaceutically acceptablecarriers are cellulose and its derivatives (such as sodiumcarboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.),gelatin, talc, solid lubricants (such as stearic acid), magnesiumstearate), calcium sulfate, vegetable oils (such as soybean oil, sesameoil, peanut oil, olive oil, etc.), polyols (such as propylene glycol,glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween®), runWet agents (such as sodium lauryl sulfate), colorants, flavoring agents,stabilizers, antioxidants, preservatives, pyrogen-free water, and thelike.

The mode of administration of the compound or pharmaceutical compositionof the present invention is not particularly limited, and representativemodes of administration include, but are not limited to, oral,intratumoral, rectal, parenteral (intravenous, intramuscular orsubcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In these solid dosage forms, the activecompound is mixed with at least one conventional inert excipient (orcarrier), such as sodium citrate or dicalcium phosphate, or mixed with:(a) a filler or compatibilizer, for example, Starch, lactose, sucrose,glucose, mannitol and silicic acid; (b) a binder such ashydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone,sucrose, and acacia; (c) a humectant such as glycerin; (d) adisintegrant, for example, Agar, calcium carbonate, potato starch ortapioca starch, alginic acid, certain complex silicates, and sodiumcarbonate; (e) a slow solvent such as paraffin; (f) an absorptionaccelerator, for example, a quaternary amine compound; (g) Wettingagents such as cetyl alcohol and glyceryl monostearate; (h) anadsorbent, for example, kaolin: and (i) a lubricant such as talc,calcium stearate, magnesium stearate, solid polyethylene glycol, sodiumlauryl sulfate, or a mixture thereof. In capsules, tablets and pills,the dosage form may also contain a buffer.

Solid dosage forms such as tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other materials known in the art. They may containopacifying agents and the release of the active compound or compound insuch compositions may be released in a portion of the digestive tract ina delayed manner. Examples of embedding components that can be employedare polymeric and waxy materials. If necessary, the active compound mayalso be in microencapsulated form with one or more of theabove-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups or elixirs. Inaddition to the active compound, the liquid dosage form may containinert diluents conventionally employed in the art, such as water orother solvents, solubilizers and emulsifiers, for example, ethanol,isopropanol, ethyl carbonate, ethyl acetate, propylene glycol,1,3-butanediol, dimethylformamide and oils, especially cottonseed oil,peanut oil, corn germ oil, olive oil, castor oil and sesame oil or amixture of these substances.

In addition to these inert diluents, the compositions may containadjuvants such as wetting agents, emulsifying and suspending agents,sweetening agents, flavoring agents and perfumes.

In addition to the active compound, the suspension may containsuspending agents, for example, ethoxylated isostearyl alcohol,polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminum methoxide and agar or mixtures of these and thelike.

Compositions for parenteral injection may comprise a physiologicallyacceptable sterile aqueous or nonaqueous solution, dispersion,suspension or emulsion, and a sterile powder for reconstitution into asterile injectable solution or dispersion. Suitable aqueous andnonaqueous vehicles, diluents, solvents or vehicles include water,ethanol, polyols, and suitable mixtures thereof.

Dosage forms for the compounds of the invention for topicaladministration include ointments, powders, patches, propellants andinhalants. The active ingredient is admixed under sterile conditionswith a physiologically acceptable carrier and any preservatives,buffers, or, if necessary, propellants.

The compounds of the invention may be administered alone or incombination with other pharmaceutically acceptable compounds.

When a pharmaceutical composition is used, a safe and effective amountof a compound of the invention is administered to a mammal (e.g., ahuman) in need of treatment wherein the dosage is a pharmaceuticallyeffective dosage, for a 60 kg body weight. The dose to be administeredis usually from 1 to 2000 mg, preferably from 5 to 500 mg, Of course,specific doses should also consider factors such as the route ofadministration, the health of the patient, etc., which are within theskill of the skilled physician.

The Main Advantages of the Present Invention are:

1. Provided a compound of formula I.

2. Provided a novel Syk kinase inhibitor and/or Syk-HDAC dual inhibitorand the preparation and use thereof. The inhibitor can inhibit theactivities of Syk and HDAC kinases at very low concentrations.

3. Provided a pharmaceutical composition for the treatment of diseasesassociated with the activity of Syk or HDAC kinase.

The present invention will be further illustrated below with referenceto the specific examples. It should be understood that these examplesare only to illustrate the invention but not to limit the scope of theinvention. The experimental methods with no specific conditionsdescribed in the following examples are generally performed under theconventional conditions, or according to the manufacturer'sinstructions. Unless indicated otherwise, parts and percentage arecalculated by weight.

Example 1: Preparation of Compound 1R

The trifluoroacetic acid salt of compound 1Ra (590 mg, 1.69 mmol) andacetaldehyde (1.30 g, 29.45 mmol) were dissolved in methanol (30 ml).Under stirring, sodium cyanoborohydride (678 mg, 10.79 mmol) was addedin batches, and the temperature was controlled at below 10° C. Thereaction was stirred at room temperature for 1 hour, TLC monitoringshowed that the reaction was completed, then the solvent was removedunder reduced pressure at room temperature. The residue was dispersed ina saturated sodium carbonate solution and extracted with ethyl acetate(25 ml×3). The combined organic layers were washed with brine and driedover anhydrous sodium sulfate. The crude product obtained byconcentrating the filtrate under reduced pressure was separated bysilica gel column (dichloromethane/methanol=50/1 solvent mixtureelution) to obtain a yellow solid compound 1Rb (430 mg, yield 97%). ¹HNMR (CDCl₃, 400 MHz) δ 7.80 (dd, J=9.2 Hz, 2.8 Hz, 1H), 7.65 (d, J=2.8Hz, 1H), 6.74 (d, J=9.2 Hz, 1H), 4.26 (dd, J=10.8 Hz, 2.8 Hz, 1H), 4.00(dd, J=10.8 Hz, 8.4 Hz, 1H), 3.80-3.74 (m, 1H), 3.43-3.36 (m, 1H),3.08-2.90 (m, 3H), 2.50-2.46 (m, 2H), 2.21-2.14 (m, 1H), 1.81 (dd, J=7.6Hz, 7.2 Hz, 1H), 1.13 (t, J=7.2 Hz, 3H); MS m/z 264.2 [M+H]⁺.

Compound 1Rb (408 mg, 1.55 mmol) was placed in a 50 ml single-neckedflask, dissolved in methanol (3 ml), and then Pd/C (10%, 40 mg) wasadded. The air in the bottle was replaced with hydrogen gas, and thereaction was stirred at room temperature for 1 hour under a hydrogenatmosphere (one atmosphere). TLC monitoring showed that the reaction wascompleted, the reaction mixture was filtered, and the filtrate wasconcentrated under reduced pressure to give a brown solid compound 1Re(361 mg, yield 99%). The compound was used in the next step withoutpurification.

Compound 1Rc (33 mg, 0.142 mmol) was suspended in isopropanol (2 ml),and DIPEA (55 mg, 0.426 mmol) and 6, 8-dibromo-imidazole [1,2-a]pyrazine (Compound 1a, 39 mg, 0.142 mmol) was added, then the reactionmixture was heated to 80° C. to react for 16 hours. The reaction mixturewas cooled to room temperature and concentrated under reduced pressure.The crude product was purified by preparative thin chromatographed(dichloromethane/methanol=40/1 mixed solvent elution) to obtain paleyellow solid compound 1Rd (28 mg, 46% yield). MS m/z 429.0 [M+H]⁺, 431.0[M+H]⁺.

Compound 1Rd (28 mg, 0.065 mmol), Compound 1b (21 mg, 0.085 mmol) andsodium Carbonate (21 mg, 0.196 mmol) were dissolved in 1,4-dioxane/water(2 ml/0.4 ml), and then Pd(PPh₃)₄ (8 mg, 0.007 mmol) was added. Thereaction system was replaced with argon for 3 times, then the reactionmixture was heated in a microwave reactor to 100° C. and stirred for 30minutes. The reaction liquid was cooled to room temperature, and thenpoured into water and extracted with ethyl acetate (10 ml×3). Thecombined organic layer was washed with brine, then dried over anhydroussodium sulfate and filtered. The crude product obtained by concentratingthe filtrate under reduced pressure was purified by preparativechromatography (methylene chloride/methanol=15/1 mixed solvent elution)to obtain pale yellow solid compound 1R (6 mg, yield 20%). ¹H NMR(DMSO-d₆, 400 MHz) δ 13.22 (s, 1H), 9.44 (s, 1H), 8.64 (s, 1H), 8.14 (s,1H), 8.09 (s, 1H), 7.98 (s, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.74-7.67 (m,2H), 7.63 (s, 1H), 7.57 (d, J=2.0 Hz, 1H), 6.88 (d, J=8.8 Hz, 1H), 4.27(dd, J=10.4 Hz, J=2.0 Hz, 1H), 3.94 (dd, J=9.8 Hz, 9.8 Hz, 1H), 3.72 (d,J=11.2 Hz, 1H), 3.03-2.97 (m, 2H), 2.92 (d, J=10.4 Hz, 1H), 2.67-2.58(m, 1H), 2.43-2.32 (m, 2H), 2.14-2.07 (m, 1H), 1.70 (dd, J=10.8 Hz, 10.4Hz, 1H), 1.05 (t, J=7.2 Hz, 3H); MS m/z 467.2 [M+H]⁺.

Example 2: Preparation of Compound 2R

Trifluoroacetic acid salt of compound 1Ra (150 mg, 0.43 mmol) and1-ethoxy-1-trimethylsiloxycyclopropane (234 mg, 1.34 mmol) weredissolved in methanol (6 ml), and sodium cyanoborohydride (84 mg, 1.34mmol) was added under stirring. The reaction system was heated to 65° C.and stirred for 16 hours, TLC monitoring shown that the reaction wascompleted. The reaction system was cooled to room temperature, thenpoured into saturated sodium carbonate solution (10 ml), and extractedwith ethyl acetate (15 ml×3). The combined organic layer was washed withbrine (15 ml) and dried over anhydrous sodium sulfate and filtered. Thecrude product obtained by concentrating the filtrate under reducedpressure was separated by silica gel column(dichloromethane/methanol=60/1 solvent mixture elution) to obtain ayellow solid compound 2Ra (95 mg, yield 80%). ¹H NMR (CDCl₃, 400 MHz) δ7.79 (dd, J=9.2 Hz, 2.4 Hz, 1H), 7.65 (d, J=2.8 Hz, 1H), 6.73 (d, J=9.2Hz, 1H), 4.26 (dd, J=10.8 Hz, 3.2 Hz, 1H), 3.99 (dd, J=10.8 Hz, 8.4 Hz,1H), 3.79-3.71 (m, 1H), 3.34-3.27 (m, 1H), 3.15-3.08 (m, 1H), 3.04-2.88(m, 2H), 2.47-2.41 (m, 1H), 2.09 (dd, J=10.4 Hz, 10.0 Hz, 1H), 1.71-1.65(m, 1H), 0.55-0.42 (m, 4H); MS m/z 276.2 [M+H]⁺.

Compound 2Ra (30 mg, 0.11 mmol) was placed in a 50 ml single-neckedflask, dissolved in methanol (3 ml), and then Pd/C (10%, 10 mg) wasadded. The bottle was replaced with hydrogen, and the reaction wasstirred at room temperature for 1 hour under a hydrogen atmosphere (oneatmosphere). TLC monitoring showed that the reaction was completed, thereaction mixture was filtered, and the filtrate was spin dried to give abrown solid compound 2Rb (25 mg, yield 94%. The compound was used in thenext step without purification.

Compound 2Rb (28 mg, 0.114 mmol), 6,8-dibromo-imidazole[1,2-a]pyrazine(Compound 1a, 34 mg, 0.123 mmol) and N, N-diisopropylethylamine (29 mg,0.225 mmol) were dissolved in isopropanol (1 ml). The reaction systemwas heated to 80° C. in a sealing tube and stirred for 16 hours. Thereaction liquid was cooled to room temperature, and then poured intowater (10 ml) and extracted with ethyl acetate (10 ml×3). The combinedorganic layer was washed with saturated brine (10 ml), then dried overanhydrous sodium sulfate and filtered. The crude product obtained byconcentrating the filtrate under reduced pressure was purified bypreparative chromatography (methylene chloride/methanol=50/1 mixedsolvent elution) to obtain pale gray solid compound 2Rc (40 mg, yield79%). MS m/z 441.0 [M+H]⁺, 443.0 [M+H]⁺.

Compound 2Rc (40 mg, 0.091 mmol), Compound 1b (32 mg, 0.131 mmol) andsodium carbonate (24 mg, 0.226 mmol) were dissolved in 1,4-dioxane/water(1 ml/0.2 ml), and then Pd(PPh₂)₂ (5 mg, 0.007 mmol) was added. Thereaction system was replaced with argon for 3 times, then the reactionsystem was heated in a microwave reactor to 100° C. and stirred for 2hours. The reaction liquid was cooled to room temperature, and thenpoured into water (10 ml) and extracted with ethyl acetate (10 ml×3).The combined organic layer was washed with brine (10 ml), then driedover anhydrous sodium sulfate and filtered. The crude product obtainedby concentrating the filtrate under reduced pressure was purified bypreparative chromatography (methylene chloride/methanol=20/1 mixedsolvent elution) to obtain yellow solid compound 2R (17 mg, yield 39%).¹H NMR (DMSO-d₆, 400 MHz) δ 13.22 (s, 1H), 9.45 (s, 1H), 8.83 (s, 1H),8.13 (s, 1H), 8.10-8.08 (m, 1H), 7.98 (d, J=0.8 Hz, 1H), 7.87-7.82 (m,1H), 7.73-7.67 (m, 2H), 7.63 (d, J=0.8 Hz, 1H), 7.57 (d, J=2.4 Hz, 1H),6.88 (d, J=8.8 Hz, 1H), 4.30-4.24 (m, 1H), 3.95 (dd, J=10.4 Hz, 9.2 Hz,1H), 3.75-3.67 (m, 1H), 3.06-2.90 (m, 3H), 2.59-2.37 (m, 2H), 2.04-1.98(m, 1H), 1.71-1.63 (m, 1H), 0.49-0.43 (m, 2H), 0.40-0.35 (m, 2H); MS m/z479.2 [M+H]⁺.

Example 3: Preparation of Compound 3R

Compound 1Ra (430 mg, 1.83 mmol) and N,N-diisopropylethylamine (706 mg,5.46 mmol) were dissolved in dry tetrahydrofuran (8 ml), and2,2,2-trifluoroethyl trifluoromethanesulfonate (551 mg, 2.37 mmol) wasadded under stirring. The reaction system was heated to 60° C. andstirred for 16 hours, TLC monitoring shown that the reaction wascompleted. The reaction system was poured into water, and extracted withethyl acetate (15 ml×3). The combined organic layer was washed withsaturated brine (15 ml) and dried over anhydrous sodium sulfate andfiltered. The crude product obtained by concentrating the filtrate underreduced pressure was separated by silica gel column(dichloromethane/methanol=3/1 solvent mixture elution) to obtain ayellow solid compound 3Ra (550 mg, yield 95%). ¹H NMR (CDCl₃, 400 MHz) δ7.80 (dd, J=9.2 Hz, 2.8 Hz, 1H), 7.66 (d, J=2.8 Hz, 1H), 6.75 (d, J=9.2Hz, 1H), 4.25 (dd, J=10.8 Hz, 2.8 Hz, 1H), 3.99 (dd, J=11.2 Hz, 8.4 Hz,1H), 3.81-3.75 (m, 1H), 3.48-3.41 (m, 1H), 3.10-2.95 (m, 5H), 2.70-2.63(m, 1H), 2.31 (dd, J=10.8 Hz, 10.8 Hz, 1H); MS m/z 318.2 [M+H]⁺.

Compound 3Ra (300 mg, 0.95 mmol) was placed in a 50 ml single-neckedflask, dissolved in methanol (15 ml), and then Pd/C (10%, 50 mg) wasadded. The bottle was replaced with hydrogen, and the reaction wasstirred at room temperature for 1 hour under a hydrogen atmosphere (oneatmosphere). TLC monitoring showed that the reaction was completed, thereaction mixture was filtered, and the filtrate was concentrated underreduced pressure to give a brown solid compound 3Rb (250 mg, yield 92%).

Compound 3Rb (35 mg, 0.122 mmol), 6,8-dibromo-imidazole[1,2-a]pyrazine(Compound 1a, 41 mg, 0.148 mmol) and N, N-diisopropylethylamine (31 mg,0.240 mmol) were dissolved in isopropanol (1 ml). The reaction systemwas heated to 80° C. in a sealing tube and stirred for 16 hours. Thereaction liquid was cooled to room temperature, and then poured intowater and extracted with ethyl acetate (10 ml×3). The combined organiclayer was washed with brine (10 ml), then dried over anhydrous sodiumsulfate and filtered. The crude product obtained by concentrating thefiltrate under reduced pressure was purified by preparativechromatography (methylene chloride/methanol=70/1 mixed solvent elution)to obtain gray solid compound 3Rc (45 mg, yield 76%). MS m/z 483.0[M+H]⁺, 485.0 [M+H]⁺.

Compound 3Rc (45 mg, 0.093 mmol), 1b (34 mg, 0.139 mmol) and sodiumcarbonate (24 mg, 0.236 mmol) were dissolved in 1,4-dioxane/water (1ml/0.2 ml), and then Pd(PPh₂)₂ (5 mg, 0.007 mmol) was added. Thereaction system was replaced with argon for 3 times, then the reactionsystem was heated in a sealing tube to 100° C. and stirred for 16 hours.The reaction liquid was cooled to room temperature, and then poured intowater (10 ml) and extracted with ethyl acetate (10 ml×3). The combinedorganic layer was washed with saturated brine (10 ml), then dried overanhydrous sodium sulfate and filtered. The crude product obtained byconcentrating the filtrate under reduced pressure was purified bypreparative chromatography (methylene chloride/methanol=15/1 mixedsolvent elution) to obtain yellow solid compound 3R (25 mg, yield 52%).¹H NMR (DMSO-d₆, 400 MHz) δ 13.22 (s, 1H), 9.47 (s, 1H), 8.64 (s, 1H),8.13 (s, 1H), 8.09 (s, 1H), 7.98 (s, 1H), 7.87-7.82 (m, 1H), 7.73-7.67(m, 2H), 7.63 (d, J=0.8 Hz, 1H), 7.59 (d, J=2.4 Hz, 1H), 6.90 (d, J=8.8Hz, 1H), 4.30-4.22 (m, 1H), 3.97-3.89 (m, 1H), 3.77-3.70 (m, 1H),3.33-3.24 (m, 2H), 3.08-2.94 (m, 3H), 2.70-2.56 (m, 2H), 2.24-2.18 (m,1H); MS m/z 521.2 [M+H]⁺.

Example 4: Preparation of Compound 4R

Compound 1Ra (970 mg, 4.12 mmol) and N-methyl-4-piperidone (933 mg, 8.24mmol were dissolved in methanol (35 ml), and acetic acid (247 mg, 4.12mmol) was added under stirring, and then sodium cyanoborohydride (776mg, 12.35 mmol) was added in batches. The temperature was controlledbelow 10° C. The reaction system was stirred at room temperature for 16hour, TLC monitoring showed that the reaction was completed, then thesolvent was removed under reduced pressure at room temperature. Theresidue was dispersed in a saturated sodium carbonate solution andextracted with ethyl acetate (25 ml×3). The combined organic layer waswashed with saturated brine (20 ml) and dried over anhydrous sodiumsulfate. The residue obtained by concentrating the filtrate underreduced pressure was separated by silica gel column(dichloromethane/methanol=20/1 solvent mixture elution) to obtain yellowsolid compound 4Ra (860 mg, yield 63%). MS m/z 333.2 [M+H]⁺.

Compound 4Ra (373 mg, 1.12 mmol) was placed in a 50 ml single-neckedflask, dissolved in methanol (7 ml), and then Pd/C (10%, 40 mg) wasadded. The bottle was replaced with hydrogen, and the reaction wasstirred at room temperature for 2 hours under a hydrogen atmosphere. TLCmonitoring showed that the reaction was completed, then the reactionsolution was filtered, and the filtrate was concentrated under reducedpressure to give crude product, which was separated by silica gel column(dichloromethane/methanol/aqueous ammonia=10/1/0.25 mixed solventelution) to provide brown solid compound 4Rb (150 mg, yield 44%). MS m/z303.3 [M+H]⁺.

Compound 4Rb (34 mg, 0.113 mmol) was suspended in isopropanol (2 ml),and DIPEA (29 mg, 0.225 mmol) and 6,8-dibromo-imidazole [1,2-a] pyrazine(31 mg, 0.113 mmol) was added, then the reaction mixture was heated to80° C. to react for 4 hours. The reaction mixture was cooled to roomtemperature. The residue obtained by concentration under reducedpressure was purified by preparative thin chromatographe(dichloromethane/methanol=15/1 mixed solvent elution) to obtain brownsolid compound 4Rc (28 mg, 50% yield). MS m/z 498.0 [M+H]⁺, 500.0[M+H]⁺.

Compound 4Rc (28 mg, 0.056 mmol), compound 1b (27 mg, 0.112 mmol) andsodium carbonate (18 mg, 0.169 mmol) were dissolved in 1,4-dioxane/water(1 ml/0.2 ml), and then Pd(PPh₃)₄ was added (6 mg, 0.006 mmol). Thereaction system was replaced with argon for 3 times, then the reactionmixture was heated in a microwave reactor to 135° C. and stirred for 1hour. The reaction liquid was cooled to room temperature, and thenpoured into water (10 ml) and extracted with ethyl acetate (10 ml×3).The combined organic layer was washed with saturated brine (10 ml), thendried over anhydrous sodium sulfate and filtered. The crude productobtained by concentrating the filtrate under reduced pressure waspurified by preparative chromatography (methylene chloride/methanol=5/1mixed solvent elution) to obtain pale pale yellow solid compound 4R (7mg, yield 23%). ¹H NMR (DMSO-d₆, 400 MHz) δ 13.22 (s, 1H), 9.44 (s, 1H),8.64 (s, 1H), 8.13 (s, 1H), 8.09 (s, 1H), 7.98 (d, J=0.8 Hz, 1H), 7.84(d, J=8.4 Hz, 1H), 7.73-7.67 (m, 2H), 7.63 (d, J=0.8 Hz, 1H), 7.57 (d,J=2.0 Hz, 1H), 6.87 (d, J=8.8 Hz, 1H), 4.26 (dd, J=10.4 Hz, J=2.4 Hz,1H), 3.94 (dd, J=10.4 Hz, 9.2 Hz, 1H), 3.72 (d, J=11.6 Hz, 1H),3.04-2.90 (m, 3H), 2.86-2.78 (m, 2H), 2.63-2.55 (m, 1H), 2.38-2.28 (m,2H), 2.23-2.11 (m, 4H), 1.95-1.85 (m, 2H), 1.80-1.71 (m, 2H), 1.49-1.38(m, 2H); MS m/z 536.4 [M+H]⁺.

Example 5: Preparation of Compound 5R

Compound 1Ra (600 mg, 2.55 mmol) and tetrahydropyrone (511 mg, 5.10mmol) were dissolved in methanol (25 ml), and acetic acid (153 mg, 2.55mmol) was added under stirring, and then sodium cyanoborohydride (481mg, 7.65 mmol) was added in batches. The temperature was controlledbelow 10° C. The reaction system was stirred at room temperatureovernight, TLC monitoring showed that the reaction was completed, thenthe solvent was removed under reduced pressure at room temperature. Theresidue was dispersed in a saturated sodium carbonate solution andextracted with ethyl acetate (20 ml×3). The combined organic layer waswashed with saturated brine (20 ml) and dried over anhydrous sodiumsulfate. The residue obtained by concentrating the filtrate wasseparated by silica gel column (dichloromethane/methanol=30/1 solventmixture elution) to obtain yellow solid compound 5Ra (800 mg, yield98%). MS m/z 320.2 [M+H]⁺.

Compound 4Ra (200 mg, 0.63 mmol) was placed in 50 ml single-neckedflask, dissolved in methanol (5 ml), and then Pd/C (10%, 30 mg) wasadded. The bottle was replaced with hydrogen, and the reaction wasstirred at room temperature for 1 hour under a hydrogen atmosphere. TLCmonitoring showed that the reaction was completed, then the reactionsolution was filtered, and the filtrate was concentrated under reducedpressure to give crude product, which was separated by silica gel column(dichloromethane/methanol=30/1 mixed solvent elution) to provide brownsolid compound 5Rb (80 mg, yield 44%). MS m/z 290.2 [M+H]⁺.

Compound 5Rb (30 mg, 0.104 mmol) was suspended in isopropanol (2 ml),and DIPEA (27 mg, 0.208 mmol) and 6,8-dibromo-imidazole [1,2-a] pyrazine(29 mg, 0.104 mmol) was added, then the reaction mixture was heated to80° C. to react for 16 hours. The reaction mixture was cooled to roomtemperature. The residue obtained by concentration under reducedpressure was purified by preparative thin chromatography(dichloromethane/methanol=15/1 mixed solvent elution) to obtain paleyellow solid compound 5Rc (43 mg, 86% yield). MS m/z 485.1 [M+H]⁺, 487.1[M+H]⁺.

Compound 5Rc (43 mg, 0.089 mmol), compound 1b (43 mg, 0.178 mmol) andsodium carbonate (28 mg, 0.267 mmol) were dissolved in 1,4-dioxane/water(1 ml/0.2 ml), and then Pd(PPh₃)₄ was added (10 mg, 0.009 mmol). Thereaction system was replaced with argon for 3 times, then the reactionmixture was heated in a microwave reactor to 135° C. and stirred for 1hour. The reaction liquid was cooled to room temperature, and thenpoured into water (10 ml) and extracted with ethyl acetate (10 ml×3).The combined organic layer was washed with saturated brine (10 ml), thendried over anhydrous sodium sulfate and filtered.

The residue obtained by concentrating the filtrate under reducedpressure was purified by preparative chromatography (methylenechloride/methanol=15/1 mixed solvent elution) to obtain pale yellowsolid compound 5R (21 mg, yield 45%). ¹H NMR (DMSO-d₆, 400 MHz) δ 13.22(s, 1H), 9.45 (s, 1H), 8.64 (s, 1H), 8.14 (s, 1H), 8.09 (s, 1H), 7.98(d, J=0.8 Hz, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.73-7.66 (m, 2H), 7.63 (d,J=0.8 Hz, 1H), 7.57 (d, J=2.4 Hz, 1H), 6.88 (d, J=9.2 Hz, 1H), 4.27 (dd,J=10.4 Hz, 2.0 Hz, 1H), 3.98-3.87 (m, 3H), 3.73 (d, J=11.2 Hz, 1H),3.30-3.24 (m, 2H), 3.06-2.91 (m, 3H), 2.63-2.55 (m, 1H), 2.47-2.39 (m,1H), 2.36-2.28 (m, 1H), 1.90 (dd, J=11.2 Hz, 11.2 Hz, 1H), 1.77-1.72 (m,2H), 1.49-1.37 (m, 2H); MS m/z 523.2 [M+H]⁺.

Example 6: Preparation of Compound 6

Compound 6a (300 mg, 1.45 mmol) and 6b (356 mg, 2.17 mmol) weredissolved in tetrahydrofuran (10 ml). The reaction was stirred at roomtemperature 4 hours, then NaBH₃CN (273 mg, 4.34 mmol) was added to thereaction system in batches. The reaction system was stirred at roomtemperature in sealed state for 16 hours. LCMS monitored that thereaction was completed, then the reaction mixture was poured into water(30 ml), and extracted with ethyl acetate (15 ml×3), and then washedwith saturated brine (10 ml×2). The ethyle acetate layer was dried overanhydrous sodium sulfate and filtered, and the residue obtained byconcentrating the filtrate under reduced pressure was purified by columnchromatography (petroleum ether/ethyl acetate=10/1 to 4/1) to give ayellow solid compound 6c (170 mg, yield 33%). ¹H NMR (CDCl₃, 400 MHz) δ8.14-8.10 (m, 1H), 8.03-8.00 (m, 1H), 7.45-7.42 (m, 2H), 6.81 (dd, J=7.2Hz, 2.0 Hz, 2H), 3.92 (s, 3H), 3.62 (s, 2H), 3.43 (t, J=5.2 Hz, 4H),2.60 (t, J=5.2 Hz, 4H); MS m/z 356.3 [M+H]⁺.

Compound 6c (150 mg, 0.42 mmol) was dissolved in methanol (5 ml), thenPd/C (10%, 50 mg) was carefully added to the reaction system, and thenreplaced with hydrogen for three times. The reaction solution wasstirred under room temperature to react for 16 hours. After LCMSdetected that the reaction was completed, the mixture was filteredthrough celite, washed with anhydrous methanol, and the filtrate wasconcentrated under reduced pressure to provide pale yellow solidcompound 6d (100 mg, yield 73%). MS m/z 326.3 [M+H]⁺.

Compound 1a (170 mg, 0.62 mmol), 6d (100 mg, 0.31 mmol) and DIPEA (80mg, 0.62 mmol) were added to isopropanol (4 ml). The reaction system washeated to 80° C. and stirred to react in a sealed system for 16 hours.LCMS monitored that the reaction was completed, then the reactionmixture was poured into water (10 ml), and extracted with ethyl acetate(10 ml×3), and then washed with saturated brine (5 ml×2). The ethylacetate layer was dried over anhydrous sodium sulfate and filtered, andthe residue obtained by concentrating the filtrate under reducedpressure was purified by column chromatography (petroleum ether/ethylacetate=10/1 to 2/1 solvent mixture) to give a brown solid compound 6e(150 mg, yield 47%). ¹H NMR (CDCl₃, 400 MHz) δ 8.03-7.99 (m, 3H), 7.71(d, J=8.8 Hz, 2H), 7.67 (s, 1H), 7.53 (d, J=0.8 Hz, 1H), 7.49 (d, J=0.8Hz, 1H), 7.45 (d, J=8.0 Hz, 2H), 6.97 (d, J=9.2 Hz, 2H), 3.92 (s, 3H),3.62 (s, 2H), 3.21-3.17 (m, 4H), 2.65-2.61 (m, 4H); MS m/z 521.2 [M+H]⁺;523.2 [M+H]⁺.

Compound 6e (140 mg, 0.27 mmol), 1b (132 mg, 0.54 mmol), Pd(PPh₃)₄ (31mg, 0.027 mmol) and Na₂CO₃ (67 mg, 0.54 mmol) were added intodioxane/water (4 ml/0.4 ml). The reaction solution was heated bymicrowave condition to 135° C. and stirred to react for 1 hours. LCMSmonitored that the reaction was completed, then the reaction mixture waspoured into water (10 ml), and extracted with ethyl acetate (10 ml×3),and then washed with saturated brine (5 ml×2). The ethyl acetate layerwas dried over anhydrous sodium sulfate and filtered, and the residueobtained by concentrating the filtrate under reduced pressure waspurified by preparative thin layer chromatography (petroleum ether/ethylacetate=1/1 solvent mixture) to give a brown solid compound 6f (80 mg,yield 53%). MS m/z 559.3 [M+H]⁺.

Compound 6f (35 mg, 0.063 mmol) was added to methanol (0.5 mL), and thenaqueous hydroxylamine solution (2 ml) was added into the reactionsolution. The reaction solution was heated to 30° C. and stirred toreact for 4 hours. LCMS monitored that the reaction was completed, thenthe reaction mixture was poured into water (5 ml), and extracted withethyl acetate (5 ml×3), and then washed with saturated brine (2 ml×2).The ethyl acetate layer was dried over anhydrous sodium sulfate andfiltered, and the residue obtained by concentrating the filtrate underreduced pressure was purified by preparative thin layer chromatography(CH₂Cl₂/MeOH=10/1 solvent mixture) to give a gray solid compound 6 (10mg, yield 28%). 1H NMR (DMSO-d₆, 400 MHz) δ 13.19 (s, 1H), 11.19 (s,1H), 9.52 (s, 1H), 9.02 (s, 1H), 8.67 (s, 1H), 8.18 (s, 1H), 8.09 (s,1H), 8.02-7.98 (m, 3H), 7.83 (d, J=8.8 Hz, 1H), 7.75-7.70 (m, 3H), 7.64(s, 1H), 7.43 (d, J=8.0 Hz, 2H), 6.99 (d, J=9.2 Hz, 2H), 3.59 (s, 2H),3.18-3.11 (m, 4H), 2.58-2.50 (m, 4H); MS m/z 560.3 [M+H]⁺.

Example 7: Preparation of Compound 7

Compound 6a (250 mg, 1.20 mmol), 7a (339 mg, 1.80 mmol), HATU (913 mg,2.40 mmol) and triethylamine (364 mg, 3.60 mmol) were added toN,N-dimethylformamide (10 ml), and the reaction mixture was stirred atroom temperature for 4 hours. LCMS monitored that the reaction wascompleted, then the reaction mixture was poured into water (30 ml), andextracted with ethyl acetate (15 ml×4), and then washed with saturatedbrine (20 ml). The organic phase was dried with anhydrous sodium sulfateand filtered. The filtrate was concentrated under reduced pressure toafford yellow liquid compound 7b (350 mg, yield 77%). MS m/z 378.2[M+H]⁺.

Compound 7b (340 mg, 0.90 mmol) was dissolved in methanol (10 ml), thenPd/C (100 mg) was carefully added to the reaction system, and thenreplaced with hydrogen for three times. The reaction solution wasstirred under room temperature to react for 2 hours. After LCMS detectedthat the reaction was completed, the filter cake was filtered throughcelite, washed with anhydrous methanol, and the organic layer wasconcentrated under reduced pressure to provide pale yellow solidcompound 7c (250 mg, yield 80%). MS m/z 348.3 [M+H]⁺.

Compound 1a (200 mg, 0.72 mmol), 7c (201 mg, 0.59 mmol) and DIPEA (153mg, 1.18 mmol) were added to isopropanol (8 ml). The reaction mixturewas heated to 80° C. in sealed tube and stirred to react overnight. LCMSmonitored that the reaction was completed, then the reaction mixture waspoured into water (20 ml), and extracted with ethyl acetate (15 ml×3),and then washed with saturated brine (10 ml×2). The organic layer wasdried over anhydrous sodium sulfate and filtered, and the residueobtained by concentrating the filtrate under reduced pressure waspurified by column chromatography (petroleum ether/ethyl acetate=10/1 to3/1 solvent mixture) to give a brown liquid compound 7d (200 mg, yield51%). ¹H NMR (CDCl₃, 400 MHz) δ 8.03 (s, 1H), 7.74 (d, J=9.2 Hz, 2H),7.69 (s, 1H), 7.54 (d, J=0.8 Hz, 1H), 7.50 (d, J=1.2 Hz, 1H), 6.97 (d,J=9.2 Hz, 2H), 3.81-3.76 (m, 2H), 3.67 (s, 3H), 3.66-3.60 (m, 2H),3.19-3.10 (m, 4H), 2.37 (t, J=8.0 Hz, 2H), 2.32 (t, J=7.6 Hz, 2H),1.69-1.60 (m, 2H), 1.50-1.42 (m, 4H), 1.40-1.32 (m, 2H); MS m/z 543.3[M+H]⁺, 545.3 [M+H]⁺.

Compound 7d (200 mg, 0.37 mmol), 1b (180 mg, 0.74), Pd(PPh₃)₄ (43 mg,0.037 mmol) and sodium carbonate (78 mg, 0.74 mmol) were added todioxane/water (8 ml/1 ml). The reaction mixture was heated by microwavecondition to 135° C. and stirred to react for 1 hour. LCMS monitoredthat the reaction was completed, then the reaction mixture was pouredinto water (15 ml), and extracted with ethyl acetate (10 ml×3), and thenwashed with saturated brine (10 ml×2). The organic layer was dried overanhydrous sodium sulfate and filtered, and the residue obtained byconcentrating the filtrate under reduced pressure was purified bypreparative thin layer chromatography (petroleum ether/ethyl acetate=1/1solvent mixture) to give gray liquid compound 7e (75 mg, yield 35%). MSm/z 581.2 [M+H]⁺.

Compound 7e (50 mg, 0.086 mmol) was dissolved in methanol (2 ml), andaqueous lithium hydroxide solution (1 N, 1 ml) was added dropwise, andthe reaction mixture was stirred at room temperature to react for 16hrs. LCMS monitored that the reaction was completed, then the reactionmixture was poured into water (4 ml), and extracted with ethyl acetate(5 ml×6), and then washed with saturated brine (3 ml×2). The organicphase was dried over anhydrous sodium sulfate and filtered. The filtratewas concentrated under reduced pressure to afford brown solid compound7f (50 mg). The compound was used in the next step reaction withoutfurther purification. MS m/z 567.4 [M+H]⁺.

Compound 7f (50 mg, 0.086 mmol), 7g (21 mg, 0.18 mmol), EDCI (34 mg,0.18 mmol), HOBT(24 mg, 0.18 mmol) and triethylamine (18 mg, 0.18 mmol)were added to tetrahydrofuran (3 ml). The reaction mixture was stirredat room temperature to react for 2 days. LCMS monitored that thereaction was completed, then the reaction mixture was poured into water(6 ml), and extracted with ethyl acetate (5 ml×5), and then washed withsaturated brine (10 ml×2). The organic layer was dried over anhydroussodium sulfate and filtered, and the residue obtained by concentratingthe filtrate under reduced pressure was purified by preparative thinlayer chromatography (petroleum ether/ethyl acetate=2/1 solvent mixture)to give a brown solid compound 7h (45 mg, yield of two steps was 69%).¹H NMR (DMSO-d₆, 400 MHz) δ 13.19 (s, 1H), 10.90 (s, 1H), 9.56 (s, 1H),8.67 (s, 1H), 8.19 (s, 1H), 8.09 (s, 1H), 8.04 (d, J=8.8 Hz, 2H), 7.99(d, J=1.2 Hz, 1H), 7.84 (d, J=8.8 Hz, 1H), 7.72 (dd, J=8.8 Hz, 0.8 Hz,1H), 7.64 (d, J=0.8 Hz, 1H), 7.02 (d, J=9.2 Hz, 2H), 4.82-4.78 (s, 1H),3.94-3.87 (m, 1H), 3.65-3.59 (m, 4H), 3.51-3.45 (m, 1H), 3.18-3.05 (m,4H), 2.35 (t, J=7.4 Hz, 2H), 1.98 (t, J=7.4 Hz, 2H), 1.70-1.58 (m, 3H),1.57-1.43 (m, 7H), 1.31-1.20 (m, 4H).

The compound 7h (40 mg, 0.06 mmol) was dissolved in tetrahydrofuran (1ml), and then 4N HCl dioxane solution (1 ml) was slowly added dropwiseat room temperature. The reaction mixture was stirred under roomtemperature to react for 2 hours. LCMS monitored that the reaction wascompleted, then the reaction mixture was concentrated under reducedpressure. The crude product was pulped and washed with acetonitrile anddiethyl ether. The product was dissolved in DMSO and added to deionizedwater, and lyophilized to give a brown solid compound 7 (25.6 mg, yield73%). ¹H NMR (DMSO-d₆, 400 MHz) δ 10.61 (br s, 1H), 10.37 (br s, 1H),8.88 (s, 1H), 8.27-8.20 (m, 3H), 8.19-8.09 (m, 4H), 7.91 (d, J=8.4 Hz,1H), 7.75 (d, J=8.4 Hz, 1H), 7.43-7.30 (m, 2H), 5.20-6.20 (br, 1H),3.82-3.70 (m, 4H), 3.36-3.22 (m, 4H), 2.37 (t, J=7.6 Hz, 2H), 1.95 (t,J=7.2 Hz, 2H), 1.56-1.43 (m, 4H), 1.34-1.20 (m, 4H); MS m/z 582.2[M+H]⁺.

Example 8: Preparation of Compound 8R

Compound 8Ra (200 mg, 0.60 mmol) was dissolved in dichloromethane (10ml), and trifluoroacetic acid was added to the reaction system (1.5 ml)and stirred at room temperature for 2 hours. Under 40° C., the mixturewas concentrated under reduced pressure to remove trifluoroacetic acid.The residue obtained was dissolved in dichloromethane (10 ml), and ethylsulfonyl chloride (116 mg, 0.90 mmol) and triethylamine (121 mg, 1.20mmol) was added thereto, and stirred at room temperature for 16 hours.The reaction mixture was washed with water (10 ml×2), then the organicphase was dried over anhydrous magnesium sulfate, filtered, and thefiltrate is concentrated under reduced pressure to obtain the filtrate,which was purified by normal phase silica gel column chromatography(dichloromethane/ethyl acetate=1:1) to provide yellow solid 8Rb (150 mg,yield 77%). MS m/z 328.0 [M+H]⁺.

Compound 8Rb (150 mg, 0.46 mmol) was dissolved in methanol (10 ml), andPd/C (10%, 25 mg) was subsequently added. The reaction mixture wasstirred at room temperature for 2 hours, filtered and the filtrate wascollected, and washed with dichloromethane/methanol (10:1) until theproduct was completely eluted. The combined filtrate was concentratedunder reduced pressure to give gray solid compound 8Rc (100 mg, yield73%). ¹H NMR (DMSO-d₆, 400 MHz) δ 6.61 (d, J=8.4 Hz, 1H), 6.09 (dd,J=8.4 Hz, 2.0 Hz, 1H), 6.03 (d, J=2.4 Hz, 1H), 4.56 (br s, 2H), 4.22(dd, J=10.4 Hz, 2.4 Hz, 1H), 3.82 (dd, J=10.4 Hz, 9.2 Hz, 1H), 3.70 (d,J=12.4 Hz, 1H), 3.59 (dd, J=10.0H, 10 Hz, 2H), 3.10 (q, J=7.2 Hz, 2H),3.01-2.95 (m, 1H), 2.91-2.82 (m, 1H), 2.70-2.55 (m, 2H), 1.22 (t, J=7.2Hz, 3H); MS m/z 298.0 [M+H]⁺.

Compound 1a (110 mg, 0.40 mmol), 8Rc (80 mg, 0.27 mmol) and DIPEA (70mg, 0.54 mmol) were added into isopropanol (3 ml). The reaction mixturewas heated in a sealed tube to 80° C. and stirred to react for 16 hrs.LCMS monitored that the reaction was completed, then the reactionmixture was poured into water (8 ml), and extracted with ethyl acetate(5 ml×4). The combined organic layers were washed with saturated brine(50 ml×2), dried over anhydrous sodium sulfate and filtered. The residueobtained by concentrating the filtrate was purified by preparative thinlayer chromatography (CH₂Cl₂/MeOH=20/1) to give pale yellow solidcompound 8Rd (80 mg, yield 40%). MS m/z 493.1 [M+H]⁺, 495.1 [M+H]⁺.

Compound 8Rd (20 mg, 0.038 mmol), 1b (19 mg, 0.076 mmol), Pd(dppf)Cl₂(2.8 mg, 0.0038 mmol) and potassium carbonate (11 mg, 0.076 mmol) wereadded to dioxane/water (1 ml/0.1 ml). The reaction mixture was heatedunder microwave conditions to 135° C. and stirred to react for 1 hour.Then 1b (9.3 mg, 0.038 mmol), Pd(dppf)Cl₂ (2.8 mg, 0.0038 mmol) andpotassium carbonate (5.3 mg, 0.038 mmol) were added, and the reactionwas heated under microwave conditions to 135° C. and stirred to reactfor 1 hour. LCMS monitored that the reaction was completed, then thereaction liquid was poured into water (4 ml), and extracted with ethylacetate (3 ml×5), and then washed with saturated brine (2 ml×2), driedover anhydrous sodium sulfate and filtered. The residue obtained byconcentrating the filtrate was purified by preparative thin layerchromatography (dichloromethane/ethyl acetate=2/1 to give the product,and then pulped with methanol and diethyl ether for two times to providebrown solid compound 8R (5.9 mg, yield 27%). ¹H NMR (DMSO-d₆, 400 MHz) δ13.23 (s, 1H), 9.51 (s, 1H), 8.65 (s, 1H), 8.13 (s, 1H), 8.10 (s, 1H),7.99 (s, 1H), 7.85 (d, J=8.4 Hz, 1H), 7.74-7.70 (m, 2H), 7.66-7.60 (m,2H), 6.95 (d, J=8.8 Hz, 1H), 4.39-4.34 (m, 1H), 3.99-3.89 (m, 2H),3.72-3.63 (m, 2H), 3.18-2.99 (m, 4H), 2.74-2.65 (m, 2H), 1.24 (t, J=7.2Hz, 3H). MS m/z 531.2 [M+H]⁺.

Example 9: Preparation of Compound 9R

Compound 1Ra trifluoroacetate (500 mg, 1.43 mmol) and DIPEA (554 mg,4.29 mmol) were added to dichloromethane (20 ml), and thentrifluoroacetic anhydride (601 mg, 2.86 mmol) was added slowly at roomtemperature. The reaction mixture was stirred at room temperature for 2hours. LCMS monitored that the reaction was completed, then the reactionmixture was poured into water (50 ml), and extracted withdichloromethane (20 ml×4). The combined organic phase was washed withsaturated brine (10 ml×2), dried with anhydrous sodium sulfate,filtered, the filtrate was concentrated under reduced pressure. Theresidue was separated and purified by preparative thin layerchromatography (petroleum ether/ethyl acetate=2/1) to provide yellowsolid compound 9Ra (320 mg, yield 68%). ¹H NMR (CDCl₃, 400 MHz) δ7.86-7.80 (m, 1H), 7.70 (d, J=2.4 Hz, 1H), 6.82 and 6.78 (two d, J=9.2Hz, 8.8 Hz, 1H), 4.71-4.65 and 4.60-4.55 (two m, 1H), 4.39-4.33 (m, 1H),4.19-3.90 (m, 3H), 3.52-3.36 (m, 2H), 3.19-3.00 (m, 2H), 2; MS m/z 332.2[M+H]⁺.

Compound 9Ra (200 mg, 0.60 mmol) was dissolved in methanol (6 ml), thenPd/C (10%, 100 mg) was carefully added to the reaction system andreplaced with hydrogen for three times. The reaction mixture was stirredto react at room temperature for 2 hours. After LCMS detected that thereaction was completed, the mixture was filtered through celite, and thefilter cake was washed with anhydrous methanol. The combined organiclayer was concentrated under reduced pressure to provide brown solidcompound 9Rb (170 mg, yield 93%). MS m/z 302.3 [M+H]⁺.

Compound 1a (291 mg, 1.06 mmol), 9Rb (160 mg, 0.53 mmol) and DIPEA (137mg, 1.06 mmol) were added to isopropanol (8 ml), and the reactionmixture was heated in a sealed tube to 80° C. and stirred to react for 4hrs. LCMS monitored that the reaction was completed, then the reactionmixture was poured into water (20 ml), and extracted with ethyl acetate(10 ml×3). The combined organic layer was washed with saturated brine (5ml×2), dried over anhydrous sodium sulfate and filtered. The residueobtained by concentrating the filtrate under reduced pressure waspurified by preparative thin layer chromatography(dichloromethane/methanol=10/1) to give a brown solid compound 9Rc (120mg, yield 45%). MS m/z 497.2 [M+H]⁺, 499.2 [M+H]⁺.

Compound 9Rc (27 mg, 0.054 mmol), 1b (26 mg, 0.108), Pd(dppf)Cl₂ (4 mg,0.0054 mmol) and sodium carbonate (13 mg, 0.108 mmol) were added todioxane/water (1 ml/0.1 ml). The reaction mixture was heated bymicrowave condition to 100° C. and stirred to react for 1 hour. LCMSmonitored that the reaction was completed, then the reaction mixture waspoured into water (3 ml), and extracted with ethyl acetate (2 ml×3). Thecombined organic layer was washed with saturated brine (2 ml×2), driedover anhydrous sodium sulfate and filtered. The residue obtained byconcentrating the filtrate under reduced pressure was purified bypreparative thin layer chromatography (dichloromethane/methanol=1/1) togive pale yellow solid compound 9 (2 mg, yield 7%). ¹H NMR (DMSO-d₆, 400MHz) δ 13.24 (s, 1H), 9.53 (s, 1H), 8.65 (s, 1H), 8.13 (s, 1H), 8.10 (s,1H), 7.99 (s, 1H), 7.85 (d, J=8.4 Hz, 1H), 7.76-7.68 (m, 2H), 7.66-7.61(m, 2H), 7.00-6.90 (m, 1H), 4.46-4.33 (m, 2H), 4.05-3.90 (m, 3H),3.25-3.10 (m, 2H), 2.85-2.64 (m, 2H); MS m/z 535.2 [M+H]⁺.

Example 10: Preparation of Compound 10

6-Bromoindazole (0.5 g, 2.54 mmol), bis(pinacolato)diboron (0.7 g, 2.79mmol) and potassium acetate (0.7 g, 7.62 mmol) were dissolved indimethyl sulfoxide (15 mL). After replaced with nitrogen for threetimes, Pd(dppf)Cl₂.DCM (310 mg, 0.38 mmol) was added to the system, andheated to 90° C. under nitrogen atmosphere to reflux for 16 hours. TLCmonitored that the raw material disappeared, and product was formed. Thereaction was cooled to room temperature, and ethyl acetate (25 ml) andwater (25 ml) were added to dilute the reaction solution and filtered.The filter cake was soaked with ethyl acetate (15 mL) and discarded. Thefiltrate was separated, and the combined organic layer was concentratedto dry to provide crude product. The crude product was purified bysilica gel column chromatography (ethyl acetate/hexane=1/50 to 30) togive an off-white solid compound 1b (384 mg, purity 96.8%, yield 62.0%).MS m/z 245.1 [M+H]⁺.

10a (0.2 g, 1.46 mmol), PyBOP (0.8 g, 1.60 mmol) and triethylamine (0.3mL) were dissolved in N,N-dimethylformamide (3.6 mL), after replacedwith nitrogen for 3 times, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine(0.2 g, 1.75 mmol) was added and reacted at 15 to 25° C. for 5 hours.HPLC showed that the compound 10a was consumed, water (7.2 ml) wasadded, the aqueous phase was extracted with ethyl acetate (20 ml×3), andthe combined organic phase was concentrated to dry to give the crudeproduct. The crude product was purified by silica gel columnchromatography (methanol/dichloromethanol=1/100 to 30) to give anoff-white solid 1b (213 mg, purity 91.5%, yield 61.6%). MS m/z 153.0[M-THP+H]⁺.

1a (0.5 g, 1.80 mmol), 10b (0.5 g, 1.98 mmol) were dissolved inisopropyl alcohol (12.5 ml), after replaced with nitrogen for threetimes, N,N-diisopropylethylamine (0.6 ml) was added, and the mixture washeated to 80° C. to react for 7 days. The reaction solution wasconcentrated under reduced pressure to give the crude product. The crudeproduct was purified by preparative thin layer chromatography(methanol/dichloromethane=1/30) to give an off-white solid 10c (46 mg,86.5% purity). MS m/z 432.1 [M+H]⁺.

10c (0.5 g, 1.15 mmol), 1b (0.3 g, 1.4 mmol) and potassium carbonate(0.3 g, 2.3 mmol) were dissolved in dioxane (7.5 ml) and water (2.5 ml).After replaced with nitrogen for 3 times, Pd(dppf)Cl₂.DCM (95 mg, 0.1mmol) was added, and after replaced with nitrogen for 3 times, warmed to90° C. to react for 7 hours. The reaction was cooled to roomtemperature, and ethyl acetate (50 mL) was added, filtered, and thefiltrate was concentrated to dryness.

The crude product was purified by silica gel column chromatography(methanol/dichloromethane=1/50), the resulting solid was pulped withmethanol (2 ml) to provide off-white solid 10d (120 mg, 91.6% purity).MS m/z 470.4 [M+H]⁺.

10d (0.1 g, 0.2 mmol) was dissolved in methanol (10 mL), andtrifluoroacetic acid (0.5 mL) was added and reacted at 15 to 25° C. for16 hours. The reaction mixture was concentrated to dryness under reducedpressure, methanol (2 ml) was added to pulp so as to give brown solid 10(41 mg, 95.6% purity, 50% yield). ¹H NMR (400 MHz, DMSO-d₆): δ 11.12 (s,1H), 10.02 (s, 1H), 8.82 (s, 1H), 8.26-8.29 (d, J=8.7 Hz, 2H), 8.22 (s,1H), 8.07-8.11 (d, J=15.8 Hz, 2H), 7.82 (m, 3H), 7.75 (d, J=8.3 Hz, 2H).MS m/z 386.3 [M+H]⁺.

Example 11: Preparation of Compound 11

To a 250 ml three-necked bottle, p-chloronitrobenzene (11b, 5.0 g, 31.8mmol), dimethyl sulfoxide (50.2 ml), morpholine (2.8 g, 38.2 mmol) andpotassium carbonate (6.6 g, 47.7 mmol) was added. The system wasreplaced with nitrogen for three times, and the reaction was heated to100° C. and stirred to react for 24 hours. The system was cooled to roomtemperature, and water (150 ml) was added to the reaction mixture, and alarge amount of solid was precipitated from the reaction mixture, Afterfiltration, the filter cake was added into ethyl acetate (40 mL) andheated with stirring to make the filter completely dissolved to form asolution. To the resulting solution, petroleum ether (100 ml) was slowlyadded dropwise within 30 minutes. Then the reaction system was cooled at0° C. for 1 hour for crystallization. After filtration, the filter cakewas dried in a vacuum oven at 60° C. for 6 hours to constant weight toobtain bright yellow solid material tic (5.6 g, purity 99.4%, yield84.9%). MS m/z 209.1 [M+H]⁺.

11c (0.6 g, 2.9 mmol) was dissolved in tetrahydrofuran (12.5 ml), andmethanol (12.5 ml) was added and stirred for 5 min. Ammonium chloride(1.5 g, 28.8 mmol) in water (6.5 mL) was added and the mixture wascontinued to stir for 30 min. Zinc powder (942 mg, 14.4 mmol) was addedto the system and the reaction was continued to stir at 17° C. for 3hours. The reaction liquid was gradually changed from bright yellow togray, and the reaction was monitored to have been finished by thin-platechromatography (methanol/dichloromethane=1:10). After stood for 15minutes, the reaction was filtered through celite, the cake was rinsedwith ethyl acetate (10.5 ml) and the filtrate was concentrated todryness under reduced pressure. Water was added (5.5 ml) and extractedwith ethyl acetate (10 ml×3). The combined organic phase was washed withsaturated sodium chloride solution (10.2 mL), dried and concentratedunder reduced pressure to give a pale yellow solid 11d (500 mg, 99.1%purity, yield 97.4%).

11d (0.5 g, 2.8 mmol), 1a (0.8 g, 2.8 mmol), isopropanol (5.1 ml), anddiisopropylethylamine (0.7 g, 5.6 mmol) were added to a 25 mlthree-necked flask. The system was replaced with nitrogen for threetimes under stirring. The system was refluxed for 16 hours in 100° C.oil bath. The system was cooled to 17° C., and concentrated to drynessunder reduced pressure, purified by silica gel column chromatography(ethyl acetate/n-hexane=1/10 to 1:3) to give light yellow solid Compound11e (0.7 g, 99.5% purity, yield 72.6%). MS m/z 374.0 [M+H]⁺.

11h (0.9 g, 3.7 mmol), 1-hydroxybenzotriazole (0.5 g, 3.7 mmol) andN,N-dimethylformamide (4.1 mL) were sequentially added to a 25 mLsingle-necked flask. After cooled to 0° C.,1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.8 g, 4.4mmol) and diisopropylethylamine (0.9 g, 7.4 mmol) were added, stirredfor 20 minutes and warmed to 13° C. 11g (0.4 g, 3.7 mmol) was added andthe reaction was continued to stirred. The reaction was monitored bythin layer chromatography (ethyl acetate: n-hexane=1:1) until thestarting material disappeared. After water (8.1 ml) was added to thereaction mixture, the mixture was extracted with ethyl acetate (10mL×3), the combined organic phase was washed with saturated sodiumchloride solution (10.2 ml), and concentrated to dry under reducedpressure. The crude product was purified by silica gel column product(ethyl acetate: n-hexane=4:1) to give a white solid compound 11f (1.0 g,purity 74.4%, yield 79.9%). MS m/z 257.0 [M-pinacol+H]⁺.

11e (240 mg, 0.6 mmol), 1,4-dioxane (3.6 ml), potassium carbonate (178mg, 1.3 mmol), water (1.2 ml), 11f (118 mg, 0.6 mmol) andPd(dppf)Cl₂.DCM (46 mg, 0.06 mmol) were added in a 25 ml three-neckedflask, and the system was replaced with nitrogen gas for three times.The system was heated to 90° C. so as to reflux for 16 hours. HPLCshowed that the reaction was completed, and the reaction mixture wascooled to 17° C. and concentrated to dry under reduced pressure. Thecrude product was purified by preparative thin-layer chromatography(methanol/dichloromethane=1/15, R_(f)=0.3), and then the silica gelcontaining the product band was passed through a flash column (550 ml,methanol/dichloromethane=1/10) so as to obtain an off-white solid 11 (35mg, purity 96.5%). ¹H NMR (400 MHz, DMSO-d₆) δ 9.76 (s, 1H), 9.56 (s,1H), 8.72 (s, 1H), 8.63 (s, 1H), 8.20 (d, J=7.8 Hz, 1H), 8.07-7.88 (m,4H), 7.64 (d, J=9.7 Hz, 2H), 7.24 (d, J=7.8 Hz, 1H), 6.96 (dd, J=21.3,8.1 Hz, 3H), 6.81 (d, J=7.9 Hz, 1H), 6.62 (t, J=7.6 Hz, 1H), 4.98 (s,2H), 3.71 (t, J=4.7 Hz, 4H), 2.99 (t, J=4.7 Hz, 4H). MS m/z 506.4[M+H]⁺.

Example 12: Preparation of Compound 12

Compound 12b was prepared by compound 12a according to the synthesis ofcompound 1b. The synthesis method of 12 from 11e and 12b by a multi-stepreaction can refer to the synthesis of compound 10. ¹H NMR (400 MHz,DMSO-d₆): δ 10.47 (s, 1H), 8.82 (s, 1H), 8.19 (s, 1H), 8.03-8.09 (m,5H), 7.70-7.72 (d, J=8.2 Hz, 1H), 7.50-7.53 (d, J=15.8 Hz, 1H), 7.33(br, 2H), 6.55-6.58 (d, J=15.8 Hz, 1H), 3.88 (t, 4H), 3.29 (t, 4H). MSm/z 457.6 [M+H]⁺.

Example 13: Preparation of Compound 13

11e (3.0 g, 8.1 mmol), 11h (2.4 g, 9.6 mmol), 1,4-dioxane (30 ml), water(6.1 ml), potassium carbonate (2.2 g, 16.1 mmol) and Pd(dppf) Cl₂.DCM(164 mg, 0.2 mmol) were added successively into a 100 ml three-neckedflask. The system was replaced with nitrogen gas for three times understirring, and then warmed to 90° C. to reflux for 16 hours. Afterfiltration, the filter cake was soaked with methanol (20 ml×3) anddiscarded. The combined organic phase was concentrated to dry underreduced pressure, and the resulting solid was washed pulped withn-hexane (3 mL)-dichloromethane (12 ml) under 21° C. for 2 h. Afterfiltration, the filter cake was washed with dichloromethane (12.3 mL)and dried in a blast oven for 16 hours under 40° C. to give light greensolid compound 13a (3.3 g, purity 94.2%, yield 98.8%). ¹H NMR (400 MHz,DMSO-d₆) δ 9.40 (s, 1H), 8.56 (s, 1H), 8.46 (t, J=1.8 Hz, 1H), 8.08-7.99(m, 3H), 7.97-7.80 (m, 4H), 7.61 (s, 1H), 7.36 (t, J=7.6 Hz, 1H),7.27-7.19 (m, 2H), 6.97 (d, J=8.9 Hz, 2H), 3.75 (t, J=4.7 Hz, 4H), 3.08(t, J=4.8 Hz, 4H).

13a (0.2 g, 0.5 mmol) was dissolved in dichloromethane (2.3 ml), and1-hydroxybenzotriazole (71 mg, 0.5 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (138 mg, 0.7mmol) and 4-dimethylamine pyridine (118 mg, 0.9 mmol) were addedsequentially under stirring. The system was stirred at 25° C. to reactfor 0.5 hours. 10b (114 mg, 0.5 mmol) was added and the reaction wasstirred at 25° C. to react for 16 h. The reaction was concentrated underreduced pressure to dryness, and purified the crude by silica gel column(methanol/dichloromethane=1/200 to 1/75) to give yellow solid 13b (192mg, purity 78.5%, yield 62.9%).

13b (190 mg, 0.3 mmol) was dissolved in methanol (2.1 mL) andtrifluoroacetic acid (216 mg, 1.5 mmol) was added dropwise at 25° C.After the dropwise addition was completed, the system was stirred for0.5 hours, and then warmed to 40° C. and stirred to react for 20 hours.The reaction solution was cooled to 25° C. After filtration, the filtercake was pulped with isopropanol (3.1 mL) at 85° C. for 1 hour. Thesystem was slowly nature cooled to 25° C. by constant stirring andfiltered. The filter cake was dried in a vacuum oven at 40° C. for 16hours to give a yellow solid 13 (30 mg, purity 90.1%, yield 18.2%). ¹HNMR (400 MHz, DMSO-d₆) δ 11.17 (s, 1H), 10.68 (s, 1H), 9.65 (s, 1H),8.78 (s, 1H), 8.61 (s, 1H), 8.22 (d, J=7.5 Hz, 1H), 8.12-7.60 (m, 11H),6.96 (t, J=9.8 Hz, 2H), 3.70 (d, J=9.1 Hz, 4H), 2.98 (t, J=4.4 Hz, 4H).MS m/z 550.5 [M+H]⁺.

Example 14: Preparation of Compound 14

6-Bromocarbazole (6.0 g, 30.6 mmol), methyl 7-bromoheptanoate (10.8 g,45.9 mmol), N,N-dimethylformamide (60.5 ml) and potassium carbonate(12.7 g, 91.8 mmol) were sequentially added to a 100 ml three-neckedflask. The system was heated to 100° C. and stirred to react for 6.5hours. The reaction was monitored by thin layer chromatography (ethylacetate/n-hexane=1/1, UV=254 nm) until the starting materialdisappeared. Water (120 ml) was added after the reaction mixture wascooled to 17° C., and extracted with ethyl acetate (50 ml×3). Thecombined organic phases were washed with saturated aqueous solution ofsodium chloride (120 mL×3) and dried over anhydrous sodium sulfate. Theorganic phase was concentrated under reduced pressure to give the crudeproduct, which was purified by silica gel column chromatography (ethylacetate/n-hexane=1/20 to 1/5) to give orange-red solid 14a (4.6 g,purity 92.4%, yield 42.7%). The synthesis of the compound 14 by amulti-step reaction from the compound 14a can be carried out byreferring to the synthesis steps of the above compounds 10, 11, and 12.

Compound 14: bright yellow solid (purity 91.0%). ¹H NMR (400 MHz,DMSO-d₆) δ 10.30 (s, 1H), 9.66 (s, 1H), 8.76 (s, 1H), 8.27 (s, 1H),8.13-7.94 (m, 4H), 7.90-7.69 (m, 3H), 7.08 (d, J=8.5 Hz, 2H), 4.46 (t,J=6.9 Hz, 2H), 3.80 (t, J=4.7 Hz, 4H), 3.16 (t, J=4.7 Hz, 4H), 1.90 (t,J=7.2 Hz, 4H), 1.44 (q, J=7.2 Hz, 2H), 1.36-1.21 (m, 4H). MS m/z 555.6[M+H]⁺.

Example 15: Preparation of Compound 15

The synthesis procedure of the compound 10b was referred, compound 13(0.7 g) and O-(tetrahydro-2H-pyran-2-yl)hydroxylamine was reacted at 21°C. for 5 hours to obtain a pale yellow solid compound 15a (0.6 g, purity93.6%, yield 62.7%). MS m/z 515.4 [M+H]⁺.

The synthesis process of compound 10 was referred. 15a (150 mg) andtrifluoroacetic acid were reacted at 21° C. for 24 hours to give brightyellow solid 15 (30 mg, purity 95.9%, yield 23.9%). ¹H NMR (400 MHz,DMSO-d₆) δ 11.29 (s, 1H), 9.52 (s, 1H), 9.09 (s, 1H), 8.65 (s, 1H), 8.49(s, 1H), 8.38 (s, 1H), 8.14 (d, J=7.6 Hz, 1H), 8.01 (d, J=8.6 Hz, 3H),7.75-7.53 (m, 3H), 6.97 (d, J=8.6 Hz, 2H), 3.76 (t, J=4.7 Hz, 4H), 3.09(t, J=4.8 Hz, 4H). MS m/z 431.3 [M+H]⁺.

Example 16: Preparation of Compound 16

The synthesis procedure of the compound 10b was referred, 13a (200 mg)and O-methylhydroxylamine hydrochloride was reacted to afford whitesolid compound 16 (46 mg, purity 96.9%, yield 21.4%). ¹H NMR (400 MHz,DMSO-d₆) δ 11.82 (s, 1H), 9.53 (s, 1H). 8.66 (s, 1H), 8.37 (t, J=1.8 Hz,1H), 8.17 (dt, J=7.8, 1.5 Hz, 1H), 8.05-7.97 (m, 3H), 7.72 (dt, J=7.8,1.3 Hz, 1H), 7.65 (d, J=1.1 Hz, 1H), 7.59 (t, J=7.7 Hz, 1H), 7.01-6.93(m, 2H). 3.75 (d, J=4.9 Hz, 7H), 3.12-3.05 (m, 4H). MS m/z 445.6 [M+H]⁺.

Example 17: Preparation of Compound 17

Compound 17 was prepared from compound 13a by a multi-step reaction. Thespecific experimental procedure was according to the synthesis of theabove compound 10-13. Compound 17: ¹H NMR (400 MHz, DMSO-d₆) δ 10.35 (s,1H), 9.52 (s, 1H), 8.66 (s, 1H), 8.56 (t, J=5.6 Hz, 1H), 8.42 (t, J=1.8Hz, 1H), 8.14 (dt, J=7.8, 1.5 Hz, 1H), 8.08-7.98 (m, 3H), 7.81 (dt,J=7.8, 1.5 Hz, 1H), 7.65 (d, J=1.1 Hz, 1H), 7.57 (t, J=7.8 Hz, 1H),7.02-6.94 (m, 2H), 3.75 (dd, J=5.9, 3.6 Hz, 4H), 3.13-3.04 (m, 4H). MSm/z 559.0 [M+H]⁺.

Example 18: Preparation of Compound 18

Compound 18 was prepared by a multi-step reaction from compound 18a andcompound 10c. The specific experimental procedure was according to thesynthesis of the above compound 10-13. Compound 18: ¹H NMR (400 MHz,DMSO-d₆) δ 10.35 (s, 1H), 9.37 (d, J=14.6 Hz, 1H), 8.66 (s, 1H), 8.08(d, J=26.4 Hz, 1H), 7.98-7.85 (m, 3H), 7.58 (d, J=10.2 Hz, 1H), 6.94 (d,J=8.8 Hz, 2H), 6.78 (d, J=17.7 Hz, 1H), 4.37 (s, 2H), 3.74 (dd, J=6.0,3.5 Hz, 4H), 3.61 (dt, J=8.5, 5.8 Hz, 3H), 3.07 (tt, J=5.1, 2.2 Hz, 6H),2.47-2.23 (m, 4H), 1.97 (d, J=6.8 Hz, 2H), 1.53 (dd, J=7.0, 3.6 Hz, 4H),1.24 (t, J=5.9 Hz, 7H). MS m/z 520.5 [M+H]⁺.

Example 19: Preparation of Compound 19

According to the synthetic procedure of the compound 13, 17c (100 mg)and compound 19a were reacted at room temperature for 16 hours, followedby purification to obtain bright yellow solid 19 (50 mg, purity 96.4%,yield 42.9%). ¹H NMR (400 MHz, DMSO-d₆) δ 9.51 (s, 1H), 9.08 (s, 1H),8.64 (s, 1H), 8.54 (t, J=5.6 Hz, 1H), 8.41 (t, J=1.8 Hz, 1H), 8.16-8.09(m, 1H), 8.05-7.97 (m, 3H), 7.81 (dt, J=7.8, 1.4 Hz, 1H), 7.66-7.51 (m,2H), 7.15 (dd, J=7.9, 1.5 Hz, 1H), 7.01-6.83 (m, 3H), 6.70 (dd, J=7.9,1.5 Hz, 1H), 6.56-6.48 (m, 1H), 4.80 (s, 2H), 3.75 (dd, J=6.0, 3.7 Hz,4H), 3.12-3.03 (m, 4H), 2.32 (t, J=7.4 Hz, 2H), 1.60 (d, J=10.7 Hz, 4H),1.46-1.19 (m, 6H). MS m/z 633.9 [M+H]⁺.

Example 20: Preparation of Compound 20

20a (2.1 g, 9.9 mmol), 17a (2.3 g, 11.9 mmol), potassium carbonate (4.1g, 29.7 mmol), potassium iodide (146 mg, 1.0 mmol), iodine cuprous (0.4g, 2.0 mmol), L-valine (144 mg, 1.0 mmol) and N,N-dimethylacetamide(20.2 ml) were added to a 50 ml three-necked flask successively. Thesystem was replaced with nitrogen for three times, and stirred to reactat 90° C. for 24 hours. Water (40 ml) was added, and the mixture wasextracted with ethyl acetate (30 ml×3). The combined organic phase waswashed with saturated brine (60 mL), dried, filtered, and concentratedunder reduced pressure. The crude product was purified by silica gelcolumn chromatography (hexane:ethyl acetate=10:1) to give yellow solidcompound 20b (1.0 g, purity 91.8%, yield 36.1%). MS m/z 280.9 [M+H]⁺.

20b (0.6 g, 2.1 mmol), sodium hydride (0.1 g, 3.2 mmol),N,N-dimethylformamide (6.1 ml) were added to 50 ml three-necked flask,and the system was replaced with nitrogen for three times. The reactionwas stirred for 1 hour after cooled to 0° C., and then with methyliodide (0.9 g, 6.4 mmol) was added and stirred for 1 hour. Ethyl acetate(35.5 ml) was added to the reaction mixture, and the organic phase waswashed with saturated brine (40.5 ml). The organic phase wasconcentrated to dryness and purified by silica gel column chromatography(hexane:ethyl acetate=10:1) to provide bright yellow oil 20c (1 g, yield92.1%). MS m/z 295.4 [M+H]⁺.

20c (0.4 g, 1.4 mmol), zinc powder (0.5 g, 7.1 mmol), ammonium chloride(0.7 g, 14.3 mmol), ethanol (4.2 ml) and water (0.8 ml) were added to a25 ml three-necked flask. The system was replaced with N2 for threetimes, and heated to 60° C. and stirred to react for 16 hours. Thereaction solution was suction filtered, and the filtrate wasconcentrated to remove the ethanol, and extracted with ethyl acetate (10mL) for three times. The combined organic phases were concentrated togive compound 20d (480 mg, 83.9% purity). MS m/z 265.2 [M+H]⁺.

20d (0.4 g, 1.5 mmol), 1a (0.4 g, 1.5 mmol), N,N-diisopropylethylamine(0.6 g, 4.5 mmol), isopropanol (4.1 mL) were added into reaction flask.The system was replaced with nitrogen for three times, warmed to 90° C.and stirred for 16 hours. The reaction mixture was concentrated to dryand purified by silica gel column chromatography (hexane:ethylacetate=8:1) to give compound 20e (0.6 g, purity 88.6%, and the yieldwas 82.6%). MS m/z 460.3 [M+H]⁺.

20e (0.7 g, 1.5 mmol), 20f (1.6 g, 4.6 mmol), potassium carbonate (0.4g, 3.1 mmol), Pd(dppf) Cl₂ (125 mg, 0.2 mmol), 1,4-dioxane (7.1 ml) andwater (1.4 ml) were added to pressure-resistant bottle. After the systemwas replaced with nitrogen for 3 times, the system was warmed to 110° C.and stirred to react for 16 hours. The reaction solution was filtered,concentrated to dry and purified by silica gel column chromatography(n-hexane:ethyl acetate=2:1) to gave compound 20g (0.5 g, purity 99.8%,yield 63.4%). MS m/z 498.6 [M-Boc+H]⁺.

20g (0.5 g, 0.9 mmol), sodium hydroxide (0.2 g, 4.8 mmol), methanol(48.2 ml) and water (10.2 ml) were successively added into a reactionflask, and the mixture was warmed to 40° C. and stirred to react for 16hours. The reaction mixture was adjusted to pH 3-4 with dilutehydrochloric acid and then concentrated to dry to give Compound 20h (0.3g, purity 100%). MS m/z 484.4 [M+H]⁺.

20h (0.6 g, 1.3 mmol), HOBT (0.2 g, 1.4 mmol), EDCI (0.3 g, 1.7 mmol),N,N-diisopropylethylamine (0.3 g, 2.6 mmol), H2N-OTHP (0.2 g, 1.5 mmol),and N,N-dimethylacetamide (15.2 ml) were added successively into areaction flask and stirred at room temperature for 6 hours. Water (30.5ml) was added, and the mixture was extracted with ethyl acetate (150mL×3). The combined organic phase was washed with saturated sodiumchloride (50.5 ml), and the combined organic phase was concentrated todry and purified by silica gel column chromatography(methanol:dichloromethane=1:50). The solid compound 20i (0.4 g, purity98.3% yield 56.1%) was obtained. MS m/z 583.7 [M+H]⁺.

20i (0.2 g, 0.3 mmol), methanol (4.1 ml) and trifluoroacetic acid (0.5g, 3.4 mmol) were successively added into a reaction flask and stirredat room temperature for 24 hours. The reaction solution was filtered,and the filter cake was dried in vacuo to provide compound 20 (50 mg,purity 99.5%, yield 29.2%). ¹H NMR (400 MHz, DMSO-d₆) δ 10.31 (br, 1H),9.85 (br, 1H), 8.74 (s, 1H), 8.23 (br, 2H), 8.15 (s, 1H), 8.11 (d,J=0.9, 1H), 8.05 (d, J=0.9, 2H), 7.85-7.87 (d, J=8.4 Hz, 1H), 7.74 (d,J=1.3 Hz, 1H), 7.72 (d, J=2.7 Hz, 1H), 7.31 (br, 2H), 3.45 (t, 2H), 3.11(s, 3H), 1.92 (t, J=7.2 Hz, 2H), 1.44-1.47 (m, 4H), 1.22-1.26 (m, 4H).MS m/z 499.9 [M+H]⁺.

Example 21: Preparation of Compound 21

21b (1.4 g, 9.6 mmol) was dissolved in dichloromethane (20.5 mL), and1-hydroxybenzotriazole (1.4 g, 10.6 mmol), EDCI (2.4 g, 12.5 mmol),4-dimethylaminopyridine (118 mg, 0.9 mmol), N,N-diisopropylethylamine(3.7 g, 28.9 mmol) were successively added to the system. The reactionmixture was stirred at room temperature for half an hour and then 21a(2.0 g, 9.6 mmol) was added, and the mixture was stirred at roomtemperature overnight. The reaction mixture was washed with water (10.5ml). After concentrated under reduced pressure, the organic phase waspurified by silica gel column chromatography (n-hexane/ethylacetate=10/1 mixed solvent elution) to give a bright yellow solidCompound 21c (1.7 g, yield 52.6%).

21c (1.7 g, 5.1 mmol) was placed in a 100 ml three-necked flask, andmethanol (17.5 ml), ammonium chloride (2.7 g, 50.7 mmol), zinc powder(1.6 g, 25.4 mmol), water (6.2 ml) were sequentially added into thereaction flask. The reaction system was replaced with nitrogen for threetimes, heated to 60° C., and stirred for 4.5 hours. The reaction liquidwas suction filtered to remove solid impurities, and the reaction liquidwas concentrated to remove methanol to give crude product 21d (1.6 g).

21d (1.6 g, 5.2 mmol), 1a (1.4 g, 5.2 mmol), N,N-diisopropylethylamine(2.0 g, 15.7 mmol), isopropanol (16.2 mL) were added successively to 50mL three-necked flask. The reaction system was replaced with nitrogenfor three times, warmed to 90° C., and stirred to react for 5 hours. Thereaction mixture was concentrated and purified by silica gel columnchromatography (hexane/ethyl acetate=20/1) to give a crude product 21e(1.6 g, 60.5% yield).

21e (0.5 g, 1.0 mmol), 20f (1.0 g, 3.0 mmol), Pd(dppf)Cl₂-DCM (82 mg,0.1 mmol), potassium carbonate (276 mg, 2.0 mmol), water (1.1 ml) and1,4-dioxane (5.2 ml) were successively added to sealed pot, and wasblown with nitrogen. After sealing, the reaction system was heated to110° C. and stirred to react for 6 hours. The reaction solution wassuction filtered, and the crude product was purified by silica gelcolumn chromatography (hexane/ethyl acetate=3/1) to afford compound 21f(450 mg, purity 90.9%, yield 83.6%).

21f (0.4 g, 0.8 mmol) was suspended in water (4.5 ml), and sodiumhydroxide (0.1 g, 2.5 mmol) was added. The mixture was warmed to 100°C., and the reaction was stirred for 1 hour. After cooled to roomtemperature, the pH of reaction mixture was adjusted to about 2 to 3with 37% hydrochloric acid, and the reaction mixture was concentrated todry to afford crude product 21g (0.6 g, purity 83.8%).

21g (0.6 g, 0.8 mmol) was dissolved in dichloromethane (60.5 mL) andN,N-dimethylacetamide (0.3 g, 2.5 mmol), 1-hydroxybenzene triazole (124mg, 0.9 mmol), EDCI (208 mg, 1.1 mmol) were added dropwise successivelyunder stirring. The mixture was stirred to react at room temperature for0.5 h, then H2N-OTHP (108 mg, 0.9 mmol) was added and the mixture wasstirred at room temperature overnight. Water (30.5 ml) was added to thereaction mixture, and extracted with dichloromethane (20.5 ml). Theorganic phase was concentrated and purified by silica gel columnchromatographe to give yellow-green solid 21h (160 mg, 81.0% purity,30.6% yield).

21h (160 mg, 0.2 mmol) was dissolved in methanol (3.1 mL),trifluoroacetic acid (185 mg, 1.3 mmol) was added under stirring, andreacted at room temperature for 3.5 hours and then at 40° C. for 4.5hours, then heated to 60° C. to react for 15 minutes. After the reactionmixture was cooled to room temperature, 70 mg of filter cake wasobtained. The filter cake was dissolved in N,N-dimethylformamide (1.1ml), and purified by preparative liquid phase reverse-phase columnchromatography to obtain compound 21 (20 mg, purity 97.8%, yield 14.5%).¹H NMR (400 MHz, DMSO-d₆) δ 10.35 (br, 1H), 9.61 (br, 1H), 8.71 (s, 1H),8.18 (s, 1H), 8.09 (d, J=0.9 Hz, 1H), 8.02-8.04 (m, 3H), 7.83-7.86 (d,J=8.5, 1H), 7.73 (d, J=0.9 Hz, 1H), 7.71 (d, J=1.4 Hz, 1H), 7.09 (d,J=9.1 Hz, 2H), 3.63 (4H), 3.13 (4H), 2.36 (t, J=7.4 Hz, 2H), 2.01 (t,J=7.4 Hz, 2H), 1.75 (m, 2H).

Example 22: Preparation of Compound 22

22a (4.0 g, 19.8 mmol) was dissolved in N,N-dimethylacetamide (30.5 mL),then 22b (2.2 g, 29.7 mmol), potassium carbonate (8.2 g, 59.4 mmol),potassium iodide (329 mg, 1.9 mmol), cuprous iodide (377 mg, 1.9 mmol),and L-valine (228 mg, 1.9 mmol) were added successively. The reactionsystem was replaced with nitrogen for three times, and heated to 120° C.and stirred overnight. Water (160 ml) was added to the reaction mixtureand extracted with ethyl acetate (80 mL×3), the extracted organic phasewas concentrated, and separated by silica gel column chromatography(hexane/ethyl acetate=10/1) to give 22c (2.9 g, purity 99.9%, yield76.4%).

22c (500 mg, 2.5 mmol) and 22d (5.0 ml) were added to a tank reactor,and potassium hydroxide (50 mg, 0.7 mmol) was added. The reaction systemwas blown with nitrogen, and heated to 120° C. and stirred overnight.The reaction solution was concentrated to remove the remaining 22d, andpurified by preparative thin-layer chromatography (developingsolvent:n-hexane/ethyl acetate=2/1, UV=254 nm, R_(f)=0.6) to provide 22e(400 mg, purity 73.1%, yield 52.9%).

22e (400 mg, 1.3 mmol), zinc powder (441 mg, 6.7 mmol), ammoniumchloride (722 mg, 13.5 mmol), methanol (4.1 ml) and water (1.1) wereadded to a 25 ml three-necked flask. The reaction system was replacedwith nitrogen for three times, then heated to 40° C. to react for 3hours, and warmed to 50° C. to react for 40 minutes, then warmed to 60°C. to react for 2.5 hours. The reaction solution was suction filtered,and the filtered cake was concentrated to remove methanol and water.Water (30.5 ml) was added and extracted with ethyl acetate (30 ml×3),and the combined organic phases were concentrated under reduced pressureto provide crude product 22f (320 mg, yield 89.0%).

22f (320 mg, 0.9 mmol), 1a (266 mg, 0.9 mmol), N,N-diisopropylethylamine(373 mg, 2.8 mmol), and isopropanol (3.1 mL) were added successively to50 mL three-necked flask. The reaction system was replaced with nitrogenfor three times, warmed to 90° C., and stirred to react for 5 hours. Thereaction solution was concentrated and purified by preparativethin-layer chromatography (developing solvent:n-hexane/ethylacetate=1/1, UV=254 nm, R_(f)=0.4) to provide compound 22e (250 mg,purity 97.6%, yield 56.4%).

22g (50 mg, 0.1 mmol), 20f (112 mg, 0.3 mmol), Pd(dppf)Cl₂-DCM (9 mg,0.01 mmol), potassium carbonate (30 mg, 0.2 mmol), water (0.4 ml) and1,4-dioxane (2.1 ml) were successively added to sealed pot, and wasblown with nitrogen. After sealing, the reaction system was heated to110° C. and stirred to react for 6 hours. The reaction solution wassuction filtered, and the crude product obtained by concentrating thefiltrate was purified by preparative thin-layer chromatography(developing solvent=dichloromethane/methanol=20/1, UV=254 nm, R_(f)=0.4,R_(f)=0.1) to provide 22i (24 mg, purity 90.9%).

22i (24 mg, 0.05 mmol) was dissolved in dichloromethane (5.1 mL), andN,N-dimethylacetamide (13 mg, 0.1 mmol), 1-hydroxybenzene triazole (8mg, 0.05 mmol), EDCI (13 mg, 0.06 mmol) were added dropwise successivelyunder stirring. The mixture was stirred to react at room temperature for0.5 h, then H2N-OTHP (7 mg, 0.06 mmol) was added and the mixture wasstirred at room temperature overnight. Water (10 ml) and dichloromethane(10 ml) were added to the reaction mixture, and the organic phase wasconcentrated and purified by preparative thin-layer chromatography(developing solvent:n-hexane/ethyl acetate=1/1, UV=254 nm, R_(f)=0.4) toprovide product 22j (14 mg, purity 97.9%, yield 48.2%).

22j (14 mg, 0.02 mmol) was dissolved in methanol (3.1 mL), andtrifluoroacetic acid (14 mg, 0.1 mmol) was added under stirring, andstirred at room temperature overnight, then warmed to 40° C. to reactfor 5.5 hours. The reaction solution was filtered, and the filter cakewas dried overnight at 30° C. in a vacuum oven to give pale yellow 22(1.9 mg, purity 95.4%, yield 14.5%). MS m/z 487.1 [M+H]⁺.

Example 23: Preparation of Compound 23

22a (5.1 g, 31.7 mmol) and ethylenediamine (20.1 ml) were added to areaction flask, and the air in the system was replaced with N2 gas forthree times, and then (the mixture) heated to 120° C. to react for 2hours. The raw material was detected to been consumed by thin layerchromatography (ethyl acetate/n-hexane=1/3). The reaction was cooled toroom temperature and concentrated under reduced pressure. Water (25 mL)was then added and pulped at 100° C. for 1 hour. After filtration, thefilter cake was transferred to a watch glass and placed in a forced airoven at 60° C. for drying. After drying, a bright yellow solid 23a (5.5g, yield 95.7%) was obtained.

23a (3.6 g, 26.1 mmol), 23b (3.4 g, 26.0 mmol), and triethylamine (4.0g, 52.1 mmol) were dissolved in methanol (50.5 ml) and reacted at roomtemperature for 16 hours. The reaction was monitored by thin layerchromatography (methanol/dichloromethane=1/5) until the startingmaterial point (R_(f)=0.6) disappeared and a new spot was formed(R_(f)=0.8). The reaction mixture was concentrated under reducedpressure to give a bright yellow solid compound 23c (4.6 g, purity86.2%, yield 55.6%).

23c (4.6 g, 14.5 mmol) was dissolved in N,N-dimethylformamide (50 mL) toreduce the system temperature to below 0° C. Sodium hydride (1.7 g, 43.5mmol) was slowly added portionwise, during which the temperature waskept below 0° C., and then stirred for 1 hour to react after theaddition. Methyl iodide (12.5 g, 87.0 mmol) was added slowly, and thereaction was carried out at 0° C. for 2 hours. The raw material wasdetected to been consumed by thin layer chromatography (ethylacetate/n-hexane=1/1). Water (50.1 ml) was added to the reactionmixture, and extracted with ethyl acetate (50 mL×3). The combinedorganic phase was dried, filtered, and concentrated under reducedpressure. The crude product was purified by silica gel columnchromatography (hexane:ethyl acetate=8:1 solvent mixture elution) togive yellow solid compound 23d (1.9 g, purity 88.5%, yield 41.5%). MSm/z 346.3 [M+H]⁺.

23d (1.9 g, 5.5 mmol), ammonium chloride (2.9 g, 55.0 mmol), zinc powder(1.8 g, 27.5 mmol), and water (4.1 ml) were dissolved in methanol (19.2ml). The reaction system was replaced with nitrogen for three times andheated to 50° C. to react for 4 hours. The reaction mixture was filteredafter thin layer chromatography (ethyl acetate/n-hexane=1/1) monitorshowed that the starting material was consumed. The filtrate was washedwith methanol (20.2 ml), and the organic filtrate was combined andconcentrated to dryness to give solid compound 23e (810 mg, yield46.6%).

23e (810 mg, 2.5 mmol), 1a (712 mg, 2.5 mmol), N,N-diisopropylethylamine(996 mg, 7.7 mmol) were dissolved in isopropyl alcohol (8.1 mL). Thereaction system was replaced with nitrogen for three times, and warmedto 90° C. and stirred to react overnight. The raw material was detectedto been consumed by thin layer chromatography (ethylacetate/n-hexane=1/1). The reaction was concentrated under reducedpressure, the obtained crude product was purified by silica gel columnchromatography (hexane/ethyl acetate=5/1 mixed solvent elution) toprovide yellow solid compound 23f (908 mg, purity 96.0%, yield 69.1%).

23f (300 mg, 0.6 mmol), C62-9 (606 mg, 1.7 mmol), sodium hydroxide (188mg, 4.7 mmol), Pd(dppf)Cl₂.DCM (144 mg, 0.2 mmol) were dissolved in1,4-dioxane (3.2 ml). The reaction system was replaced with nitrogen forthree times, and the mixture was warmed to 110° C. and stirred to reactovernight. The reactant was cooled to room temperature, and dilutehydrochloric acid was added to the reaction mixture to adjust the pH to4 to 5. The reaction mixture became cloudy and yellow solid was formed.Ethyl acetate (10 ml) was added to the mixture and stirred for 0.5 hr.Filtered to give a crude yellow solid, and purified by preparative thinlayer chromatography to provide yellow solid 23g (270 mg, yield 86.1%).MS m/z 535.6 [M+H]⁺.

23g (150 mg, 0.3 mmol), 1-hydroxybenzotriazole (42 mg, 0.3 mmol),carbodiimide hydrochloride (70 mg, 0.4 mmol), N,N-diisopropyl ethylamine(73 mg, 0.5 mmol) were dissolved in N,N-dimethylformamide (1.5 ml),stirred at room temperature for 30 min, and NH₂ OTHP (45.7 mg, 0.4 mmol)was added and stirred to react overnight. The reaction was cooled toroom temperature, and the crude product was purified by preparativethin-layer chromatography to provide green solid compound 23h (62 mg,88.6% purity, 41.4% yield). MS m/z 634.7 [M-Boc+H]⁺.

23h (62 mg, 0.1 mmol) was dissolved in methanol (15.2 ml),trifluoroacetic acid (177 mg, 1.5 mmol) was added, and the system waswarmed to 60° C. to react for 4 hours. The insolubles appeared in thereaction mixture was the product. Thin layer chromatography(dichloromethane:methanol=1:10) monitored that the starting material wasconsumed. After filtration, the filter cake was dried in a vacuum ovenat 40° C. to obtain bright yellow solid 23 (20 mg, purity 95.4%, yield35.5%). ¹H NMR (400 MHz, DMSO-d₆) δ 13.19 (s, 1H), 11.06 (s, 1H), 9.45(s, 1H), 8.68 (d, J=14.9 Hz, 3H), 8.16 (s, 1H), 8.10 (s, 1H), 8.01 (s,1H), 7.95 (d, J=8.5 Hz, 2H), 7.85 (d, J=8.5 Hz, 1H), 7.75-7.68 (m, 2H),6.88 (d, J=8.1 Hz, 2H), 3.82 (t, J=6.7 Hz, 2H), 3.60 (t, J=6.7 Hz, 2H),2.97 (s, 3H). MS m/z 550.5 [M+H]⁺.

Example 24

1. Syk Kinase Activity Inhibitory Experiment

Measure of SYK protein kinase activity was carried out by using theCaliper mobility shift assay. The compound was dissolved with DMSO anddiluted with kinase buffer (20 mM HEPES pH 7.5, 0.01% Triton X-100, 5 mMMgCl₂, 1 mM MnCl₂, 2 mM DTT), and 5 μl of 5 times the finalconcentration of the compounds (10% DMSO) were added to 384 well plate.10 μl of 2.5 fold enzyme (with SYK) solution was added and incubated atroom temperature for 10 minutes, then 10 μl of 2.5 fold substrate(Peptide FAM-P22) and ATP) solution was added. The mixture was incubatedfor 30 minutes under 28° C., and 25 μl of stop buffer (100 mM HEPES pH7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50 mM EDTA) was added toterminate the reaction. Conversion rate data was read by Caliper EZReader II (Caliper Life Sciences). Conversion rate was converted toinhibition rate data (% inhibition rate=(max−conversionrate)/(max−min)*100). The max refers to the conversion rate of the DMSOcontrol, and min refers to the conversion rate of the enzyme-freecontrol. The curve was drawn with the compound concentration andinhibition rate as the horizontal and vertical coordinates, and thecurve was fitted with the XLFit excel add-in version 4.3.1 Software, andIC₅₀ was calculated.

The results indicate that the IC₅₀ of majority of the tested compoundsof the present invention was 10-1000 nM, and the IC₅₀ of preferredcompound was lower than 20 nM. The activities of some representativecompounds are shown in Table 1.

TABLE 1 Inhibition of Syk kinase activity Compound Syk (IC₅₀, nM)  1R<10  2R <10  3R <500  4R <10  5R <10 6 <10 7 <10  8R <10  9R <100 11 <500 12  <100 13  <100 14  <500 17  <1000 18  <500 19  <1000 20  <50 21 <10 22  <50 23  <50

2. HDAC-1 and HDAC-6 Activity Inhibition Experiment

HDAC activity was measured by the Synergy MX Multi-Function MicroplateReader. The compounds were dissolved with DMSO, and transferred to a 384well test plate using an Echo non-contact nanoscale sonic pipettingsystem. 15 μl of enzyme (HDAC1/HDAC6, respectively) solvent was added,and incubated at room temperature for 15 minutes, then 10 μl ofsubstrate (trypsin and Ac-peptide) solution was added. Fluorescenceintensity signal was read directly on Synergy MX (fluorescenceexcitation 355 nm, emission fluorescence 460 nm) after cultivated atroom temperature for 60 minutes. Fluorescence intensity signal wasconvented into inhibition rate data (% inhibition rate=(max−fluorescenceintensity)/(max−min)*100). The max refers to the fluorescence intensityof the DMSO control, and min refers to the fluorescence intensity of theenzyme-free control. The curve was drawn with the compound concentrationand inhibition rate as the horizontal and vertical coordinates, and thecurve was fitted with the GraphPad Prism V5.0 Software, and IC₅₀ wascalculated.

TABLE 2 HDAC activity inhibition Compound HDAC1 (IC₅₀, nM) HDAC6 (IC₅₀,nM) 6 <1000 <50 7 <30 <30 10 <2000 12 <200 13 <200 14 <20 17 <50 18 <10019 <500 20 <20 21 <100 22 <500 23 <30

All literatures mentioned in the present application are incorporatedherein by reference, as though each one is individually incorporated byreference. Additionally, it should be understood that after reading theabove content, those skilled in the art can make various changes andmodifications to the present invention. These equivalents also fallwithin the scope defined by the appended claims.

1. A compound of the following formula (I), or the optical isomers,pharmaceutically acceptable salts thereof:

wherein in the formula (I), R¹ is aryl, heteroaryl or 6-memberedmonocyclic heterocyclyl (including saturated and unsaturated); aryl,heteroaryl or monocyclic heterocyclyl herein may be optionally andindependently substituted by 1-3 substituents each independentlyselected from the group consisting of: halogen, C₁₋₄ alkyl, C₁₋₄halogenated alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₈ cycloalkyl, 3- to8-membered heterocyclyl, aryl, heteroaryl, CN, NO₂, OR⁸, SR⁸, NR⁸R⁹,C(O)R⁸, C(O)OR⁸, C(O)NR⁸R⁹, S(O)₂R⁸ and R⁷; R², R³ and R⁴ are hydrogen;U is NR⁵; where R⁵ is hydrogen; A is the group of formula (III):

wherein: “

” refers to the connection point of formula (III) to U of the formula(I); “*” indicates a chiral center; each X is independently hydrogen,halogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, 3- to 6-membered heterocyclyl, CN,OR⁸, SR⁸, NR⁸R⁹, C(O)R⁸, C(O)OR⁸, C(O)NR⁸R⁹, or S(O)₂R⁸; R⁷ is hydrogen,—(CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH),—V¹—(CH₂)_(p)—V²—V—(CH₂)_(q)C(O)NH(OH), C(O)NH(OCH₃),

J is O; G is NR¹⁰; n is 0, 1, 2, or 3; each R⁶ is hydrogen, or two R⁶connecting to the same carbon atom form carbonyl group (═O); R⁸ and R⁹are each independently hydrogen, C₁₋₄ alkyl, C₁₋₄ halogenated alkyl,C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₈ cycloalkyl, 3- to 8-memberedheterocyclyl, aryl, or heteroaryl; or R⁸ and R⁹ together with thenitrogen atom to which they are attached form a 3- to 9-metacyclic ringcomprising 1-2 N atom and 0, 1 or 2 heteroatoms selected from O or S;R¹⁰ is C₂₋₈ alkyl, C₁₋₈ haloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₈cycloalkyl, 3- to 12-membered heterocyclyl (optionally comprising 1-2heteroatoms selected from O, N, or S), aryl, heteroaryl, C(O)R⁸,C(O)OR⁸, C(O)NR⁸R⁹, S(O)₂R⁸ or (CH₂)_(p)—V—(CH₂)_(q)C(O)NH(OH), whereinR⁸ is not hydrogen, and the R⁸ in C(O)R⁸ is not methyl; V is a divalentgroup, each of p and q is independently an integer from 0 to 10, and theV is selected from the group consisting of bond, O, S, NR¹¹, OC(O),OC(O)O, NHC(O), NHC(O)NH, NHC(O)O, OC(O)NH, NHS(O)₂, C(O), C(O)O,C(O)NH, S(O), S(O)₂, S(O)₂NH, or NHS(O)₂NH, CH═CH, C≡C, CR¹²R¹³, C₃₋₈cycloalkyl, 3- to 12-member heterocyclyl, aryl or heteroaryl,

with the prerequisite that V, p and q together form a chemically stablegroup; V¹ and V² are divalent groups selected from the group consistingof bond, O, S, NR¹¹, and C(O)NH, with the prerequisite that the groupformed by V, V¹, V², p and q is chemically stable group; R¹¹ ishydrogen, C₁₋₄ alkyl, C₃₋₈ cycloalkyl, 3- to 8-membered heterocyclyl,aryl, heteroaryl, C(O)R⁸ or S(O)₂R⁸; R¹² and R¹³ are each independentlyhydrogen, halogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, 3- to 8-membered heterocyclic, OR⁸, SR⁸, NR⁸R⁹, CN, C(O)R⁸,C(O)OR⁸, C(O)NR⁸R⁹, OC(O)R⁸, NR⁸C(O)R⁹, or S(O)₂R⁸, or R¹² and R¹³together with the carbon atoms to which they are attached form 3-8membered cyclic structure containing 0, 1 or 2 heteroatoms selected fromN, O, or S; with the proviso that when R¹ does not comprise structuralunit

then A is of formula (IIIa),

wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl,aryl and heteroaryl are each optionally and independently substituted by1-3 substituents each independently selected from the group consistingof halogen, C₁₋₄ alkyl, C₁₋₄ halogenated alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₈ cycloalkyl, 3- to 8-heterocyclyl, aryl, heteroaryl, CN,NO₂, OR⁸, SR⁸, NR⁸R⁹, C(O)R⁸, C(O)OR⁸, C(O)NR⁸R⁹ or S(O)₂R⁸; unlessotherwise specified, the above aryl group is an aryl group having 6 to12 carbon atoms; and the heteroaryl group is a 5- to 15-memberedheteroaryl group.
 2. The compound of claim 1, or the optical isomers,pharmaceutically acceptable salts thereof, wherein R¹ is

wherein R⁷ is as described in claim
 1. 3. The compound of claim 1, orthe optical isomers, pharmaceutically acceptable salts thereof, whereinthe formula (III) is a structure selected from group consisting of:

wherein R¹⁰ is C₂₋₈ alkyl, C₁₋₈ halogenated alkyl, C₂₋₈ alkenyl, C₂₋₈alkynyl, C₃₋₈ cycloalkyl, 6-membered heterocyclyl (optionally comprising1-2 heteroatoms selected from O, N, S), aryl, heteroaryl, C(O)R⁸,C(O)OR⁸, C(O)NR⁸R⁹, S(O)₂R⁸, wherein R⁸ is selected from the groupconsisting of C₁₋₄ alkyl, C₁₋₄ haloalkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl,C₃₋₈ cycloalkyl, 3- to 8-membered heterocyclyl, aryl, and heteroaryl;wherein R⁸ in C(O)R⁸ is other than methyl; or R⁸ and R⁹ together withthe nitrogen atom to which they are attached form a 3- to 9-memberedring comprising 1-2 N atom and 0, 1 or 2 hetero atoms selected from O orS.
 4. The compound of claim 1, or the optical isomers, pharmaceuticallyacceptable salts thereof, wherein the formula (I) is selected from groupconsisting of:

wherein R¹⁰ is C₂₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₈ cycloalkyl,unsubstituted or C₁₋₄ alkyl substituted 6-membered heterocyclyl(optionally containing 1-2 heteroatoms selected from O, or N), C(O)R⁸,or S(O)₂R⁸; R⁸ is C₁₋₄ alkyl or C₁₋₄ haloalkyl; wherein the R⁸ in C(O)R⁸is other than methyl.
 5. The compound according to claim 1, wherein thecompound is selected from the group consisting of:


6. A method for treating a disease associated with Syk and/or HDACkinase activity or expression amount, comprising the step:administrating the compound (I) of claim 1, or the optical isomers,pharmaceutically acceptable salts thereof to a subject in need thereof,wherein the disease is selected from the group consisting of lymphoma,lymphocytic leukemia, cutaneous T-cell lymphoma, rectal cancer, breastcancer, stomach cancer, pancreatic cancer, liver cancer, lung cancer,head and neck cancer, kidney cancer, colon cancer, ovarian cancer,prostate cancer, multiple sclerosis, immunity diseases, allergicdiseases, atherosclerosis, gastrointestinal disorders, idiopathicthrombocytopenic purpura, systemic lupus erythematosus, Alzheimer'sdisease, stroke and coronary artery disease, Wiskott-Aldrich syndrome,myelofibrosis, and AIDS.
 7. A pharmaceutical composition, comprising:(i) an effective amount of a compound of formula (I) according to claim1, or the optical isomers, pharmaceutically acceptable salts thereof;and (ii) pharmaceutically acceptable carriers.
 8. A method for preparingthe compound of claim 1, comprising the following steps:

(1) in an inert solvent, compound Ia reacts with A-NH₂ to providecompound Ib; (2) in an inert solvent, compound Ib reacts with compoundR¹B(OH)₂ to obtain compounds of formula I;

wherein each group is defined as in claim 1.